Anti-pd-1 antibodies, compositions comprising anti-pd-1 antibodies and methods of using anti-pd-1 antibodies

ABSTRACT

Provided herein are antibodies that selectively bind to PD-1 and its isoforms and homologs, and compositions comprising the antibodies. Also provided are methods of using the antibodies, such as therapeutic and diagnostic methods.

FIELD

Provided herein are antibodies with binding specificity for PD-1 andcompositions comprising the antibodies, including pharmaceuticalcompositions, diagnostic compositions and kits. Also provided aremethods of using anti-PD-1 antibodies for therapeutic and diagnosticpurposes.

BACKGROUND

Programmed cell death protein 1 (PD-1, also known as CD279) is a cellsurface protein molecule that belongs to the immunoglobulin superfamily.It is expressed on T and B lymphocytes and macrophages, and plays a rolein cell fate and differentiation. See Ishida et al., EMBO J., 1992,11:3887-3895, incorporated by reference in its entirety. Activation ofPD-1 is thought to negatively regulate the immune response. See Blank etal., Cancer Immunol. Immunother., 2007, 56:739-745; and Freeman et al.,J. Exp. Med., 2000, 192:1027-1034, each of which is incorporated byreference in its entirety.

PD-1 has two known ligands, PD-L1 and PD-L2, which are both members ofthe B7 family. See Freeman et al., supra; and Latchman et al., Nat.Immunol., 2001, 2:261-268, each of which is incorporated by reference inits entirety. The interaction between PD-1 and these ligands is thoughtto play a role in a variety of diseases, including cancer (see Ribas andTumeh, Clin. Cancer Res., 2014, Jun. 26, PMID: 24970841 [Epub ahead ofprint]), autoimmune disease (see Dai et al., Cell Immunol., 2014,290:72-79), and infection (see Day et al., Nature, 2006, 443:350-354).Each of the references cited in the preceding sentence is incorporatedby reference in its entirety. In particular, the engagement of PD-1 byone of its ligands is thought to inhibit T-cell effector functions in anantigen-specific manner.

In view of the role of PD-1 in multiple disease processes, there is aneed for improved methods of modulating the interaction of PD-1 with itsligands and the downstream signaling processes activated by PD-1.Moreover, given the role of PD-1 in several diseases, there is also aneed for therapeutics that specifically target cells and tissues thatexpress PD-1.

SUMMARY

Provided herein are antibodies that selectively bind PD-1. In someembodiments, the antibodies bind human PD-1. In some embodiments, theantibodies also bind homologs of human PD-1. In some aspects, thehomolog is a cynomolgus monkey homolog. In some aspects, the homolog isa murine homolog. In some embodiments, the antibodies bind to humanPD-1, a cynomolgus monkey homolog, and a murine homolog.

In some embodiments, the antibodies comprise at least one CDR sequencedefined by a consensus sequence provided in this disclosure. In someembodiments, the antibodies comprise an illustrative CDR, V_(H), orV_(L) sequence provided in this disclosure, or a variant thereof. Insome aspects, the variant is a variant with one or more conservativeamino acid substitutions.

Also provided are compositions and kits comprising the antibodies. Insome embodiments, the compositions are pharmaceutical compositions. Anysuitable pharmaceutical composition may be used. In some embodiments,the pharmaceutical composition is a composition for parenteraladministration.

This disclosure also provides methods of using the anti-PD-1 antibodiesprovided herein. In some embodiments, the method is a method oftreatment. In some embodiments, the method is a diagnostic method. Insome embodiments, the method is an analytical method. In someembodiments, the method is a method of purifying and/or quantifyingPD-1.

In some embodiments, the antibodies are used to treat a disease orcondition. In some aspects, the disease or condition is selected from acancer, autoimmune disease, and infection.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a comparison of the Kabat and Chothia numbering systemsfor CDR-H1. Adapted from Martin A. C. R. (2010). Protein Sequence andStructure Analysis of Antibody Variable Domains. In R. Kontermann & S.Dübel (Eds.), Antibody Engineering vol. 2 (pp. 33-51). Springer-Verlag,Berlin Heidelberg.

FIG. 2 provides a chart of tumor volume over 17-days of treatment withvarious anti-PD-1 antibodies, as described in Example 15.

FIG. 3 provides a chart of interferon gamma (IFN-g) secretion in acytomegalovirus (CMV) recall assay, as described in Example 16.

FIG. 4 provides a chart of interferon gamma (IFN-g) secretion in a mixedlymphocyte response (MLR) assay, as described in Example 17.

FIG. 5 provides a chart of mouse survival in a model of graft versushost disease, as described in Example 18.

DETAILED DESCRIPTION 1. Definitions

Unless otherwise defined, all terms of art, notations and otherscientific terminology used herein are intended to have the meaningscommonly understood by those of skill in the art to which this inventionpertains. In some cases, terms with commonly understood meanings aredefined herein for clarity and/or for ready reference, and the inclusionof such definitions herein should not necessarily be construed torepresent a difference over what is generally understood in the art. Thetechniques and procedures described or referenced herein are generallywell understood and commonly employed using conventional methodologiesby those skilled in the art, such as, for example, the widely utilizedmolecular cloning methodologies described in Sambrook et al., MolecularCloning: A Laboratory Manual 2nd ed. (1989) Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y. As appropriate, proceduresinvolving the use of commercially available kits and reagents aregenerally carried out in accordance with manufacturer defined protocolsand/or parameters unless otherwise noted.

As used herein, the singular forms “a,” “an,” and “the” include theplural referents unless the context clearly indicates otherwise.

The term “about” indicates and encompasses an indicated value and arange above and below that value. In certain embodiments, the term“about” indicates the designated value±10%, ±5%, or ±1%. In certainembodiments, the term “about” indicates the designated value±onestandard deviation of that value.

The term “combinations thereof” includes every possible combination ofelements to which the term refers to. For example, a sentence statingthat “if α₂ is A, then α₃ is not D; α₅ is not S; or α₆ is not S; orcombinations thereof” includes the following combinations when α₂ is A:(1) α₃ is not D; (2) α₅ is not S; (3) α₆ is not S; (4) α₃ is not D; asis not S; and α₆ is not S; (5) α₃ is not D and α₅ is not S; (6) α₃ isnot D and α₆ is not S; and (7) α₅ is not S and α₆ is not S.

The terms “PD-1” and “PD-1 antigen” are used interchangeably herein.Unless specified otherwise, the terms include any variants, isoforms andspecies homologs of human PD-1 that are naturally expressed by cells, orthat are expressed by cells transfected with a PD-1 gene. PD-1 proteinsinclude full-length PD-1 (e.g., human PD-1; GI: 167857792; SEQ ID NO: 1;extracellular domain: Pro21-Gln167), as well as alternative splicevariants of PD-1, such as PD-1Δex2, PD-1Δex3, PD-1Δex2,3, andPD-1Δex2,3,4. See Nielsen et al., Cellular Immunology, 2005,235:109-116, incorporated by reference in its entirety. In someembodiments, PD-1 proteins include murine PD-1 (e.g., SEQ ID NO: 299;extracellular domain: Leu25-Gln167). In some embodiments, PD-1 proteinsinclude cynomolgus PD-1 (e.g., SEQ ID NO: 300; extracellular domain:Pro21-Gln167).

The term “immunoglobulin” refers to a class of structurally relatedproteins generally comprising two pairs of polypeptide chains: one pairof light (L) chains and one pair of heavy (H) chains. In an “intactimmunoglobulin,” all four of these chains are interconnected bydisulfide bonds. The structure of immunoglobulins has been wellcharacterized. See, e.g., Paul, Fundamental Immunology 7th ed., Ch. 5(2013) Lippincott Williams & Wilkins, Philadelphia, Pa. Briefly, eachheavy chain typically comprises a heavy chain variable region (V_(H))and a heavy chain constant region (CH). The heavy chain constant regiontypically comprises three domains, C_(H1), C_(H2), and C_(H3). Eachlight chain typically comprises a light chain variable region (V_(L))and a light chain constant region. The light chain constant regiontypically comprises one domain, abbreviated CL.

The term “antibody” describes a type of immunoglobulin molecule and isused herein in its broadest sense. An antibody specifically includesintact antibodies (e.g., intact immunoglobulins), and antibodyfragments. Antibodies comprise at least one antigen-binding domain. Oneexample of an antigen-binding domain is an antigen binding domain formedby a V_(H)-V_(L) dimer. A “PD-1 antibody,” “anti-PD-1 antibody,” “PD-1Ab,” “PD-1-specific antibody” or “anti-PD-1 Ab” is an antibody, asdescribed herein, which binds specifically to the antigen PD-1. In someembodiments, the antibody binds the extracellular domain of PD-1.

The V_(H) and V_(L) regions may be further subdivided into regions ofhypervariability (“hypervariable regions (HVRs);” also called“complementarity determining regions” (CDRs)) interspersed with regionsthat are more conserved. The more conserved regions are called frameworkregions (FRs). Each V_(H) and V_(L) generally comprises three CDRs andfour FRs, arranged in the following order (from N-terminus toC-terminus): FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. The CDRs are involved inantigen binding, and confer antigen specificity and binding affinity tothe antibody. See Kabat et al., Sequences of Proteins of ImmunologicalInterest 5th ed. (1991) Public Health Service, National Institutes ofHealth, Bethesda, Md., incorporated by reference in its entirety.

The light chain from any vertebrate species can be assigned to one oftwo types, called kappa and lambda, based on the sequence of theconstant domain.

The heavy chain from any vertebrate species can be assigned to one offive different classes (or isotypes): IgA, IgD, IgE, IgG, and IgM. Theseclasses are also designated α, δ, ε, γ, and μ, respectively. The IgG andIgA classes are further divided into subclasses on the basis ofdifferences in sequence and function. Humans express the followingsubclasses: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

The amino acid sequence boundaries of a CDR can be determined by one ofskill in the art using any of a number of known numbering schemes,including those described by Kabat et al., supra (“Kabat” numberingscheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (“Chothia”numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732-745(“Contact” numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003,27:55-77 (“IMGT” numbering scheme); and Honegge and Plückthun, J. Mol.Biol., 2001, 309:657-70 (“AHo” numbering scheme), each of which isincorporated by reference in its entirety.

Table 1 provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1,CDR-H2, and CDR-H3 as identified by the Kabat and Chothia schemes. ForCDR-H1, residue numbering is provided using both the Kabat and Chothianumbering schemes. FIG. 1 provides a comparison of the Kabat and Chothianumbering schemes for CDR-H1. See Martin (2010), supra.

Unless otherwise specified, the numbering scheme used for identificationof a particular CDR herein is the Kabat/Chothia numbering scheme. Wherethe residues encompassed by these two numbering schemes diverge, thenumbering scheme is specified as either Kabat or Chothia.

TABLE 1 Residues in CDRs according to Kabat and Chothia numberingschemes. CDR Kabat Chothia L1 L24-L34 L24-L34 L2 L50-L56 L50-L56 L3L89-L97 L89-L97 H1 (Kabat Numbering) H31-H35B H26-H32 or H34* H1(Chothia Numbering) H31-H35 H26-H32 H2 H50-H65 H52-H56 H3 H95-H102H95-H102 *The C-terminus of CDR-H1, when numbered using the Kabatnumbering convention, varies between H32 and H34, depending on thelength of the CDR, as illustrated in FIG. 1.

The “EU numbering scheme” is generally used when referring to a residuein an antibody heavy chain constant region (e.g., as reported in Kabatet al., supra). Unless stated otherwise, the EU numbering scheme is usedto refer to residues in antibody heavy chain constant regions describedherein.

An “antibody fragment” comprises a portion of an intact antibody, suchas the antigen binding or variable region of an intact antibody.Antibody fragments include, for example, Fv fragments, Fab fragments,F(ab′)₂ fragments, Fab′ fragments, scFv (sFv) fragments, and scFv-Fcfragments.

“Fv” fragments comprise a non-covalently-linked dimer of one heavy chainvariable domain and one light chain variable domain.

“Fab” fragments comprise, in addition to the heavy and light chainvariable domains, the constant domain of the light chain and the firstconstant domain (Cm) of the heavy chain. Fab fragments may be generated,for example, by papain digestion of a full-length antibody.

“F(ab′)₂” fragments contain two Fab′ fragments joined, near the hingeregion, by disulfide bonds. F(ab′)₂ fragments may be generated, forexample, by pepsin digestion of an intact antibody. The F(ab′) fragmentscan be dissociated, for example, by treatment with ß-mercaptoethanol.

“Single-chain Fv” or “sFv” or “scFv” antibody fragments comprise a VIIdomain and a V_(L) domain in a single polypeptide chain. The V_(H) andV_(L) are generally linked by a peptide linker. See Plückthun A. (1994).Antibodies from Escherichia coli. In Rosenberg M. & Moore G. P. (Eds.),The Pharmacology of Monoclonal Antibodies vol. 113 (pp. 269-315).Springer-Verlag, New York, incorporated by reference in its entirety.“scFv-Fc” fragments comprise an scFv attached to an Fc domain. Forexample, an Fc domain may be attached to the C-terminal of the scFv. TheFc domain may follow the V_(H) or V_(L), depending on the orientation ofthe variable domains in the scFv (i.e., V_(H)-V_(L) or V_(L)-V_(H)). Anysuitable Fc domain known in the art or described herein may be used. Insome cases, the Fc domain is an IgG1 Fc domain (e.g., SEQ ID NO: 295).In some embodiments, the linker is (G₄S)₃ (see SEQ ID NO: 298).

The term “monoclonal antibody” refers to an antibody from a populationof substantially homogeneous antibodies. A population of substantiallyhomogeneous antibodies comprises antibodies that are substantiallysimilar and that bind the same epitope(s), except for variants that maynormally arise during production of the monoclonal antibody. Suchvariants are generally present in only minor amounts. A monoclonalantibody is typically obtained by a process that includes the selectionof a single antibody from a plurality of antibodies. For example, theselection process can be the selection of a unique clone from aplurality of clones, such as a pool of hybridoma clones, phage clones,yeast clones, bacterial clones, or other recombinant DNA clones. Theselected antibody can be further altered, for example, to improveaffinity for the target (“affinity maturation”), to humanize theantibody, to improve its production in cell culture, and/or to reduceits immunogenicity in a subject.

The term “chimeric antibody” refers to an antibody in which a portion ofthe heavy and/or light chain is derived from a particular source orspecies, while the remainder of the heavy and/or light chain is derivedfrom a different source or species.

“Humanized” forms of non-human antibodies are chimeric antibodies thatcontain minimal sequence derived from the non-human antibody. Ahumanized antibody is generally a human immunoglobulin (recipientantibody) in which residues from one or more CDRs are replaced byresidues from one or more CDRs of a non-human antibody (donor antibody).The donor antibody can be any suitable non-human antibody, such as amouse, rat, rabbit, chicken, or non-human primate antibody having adesired specificity, affinity, or biological effect. In some instances,selected framework region residues of the recipient antibody arereplaced by the corresponding framework region residues from the donorantibody. Humanized antibodies may also comprise residues that are notfound in either the recipient antibody or the donor antibody. Suchmodifications may be made to further refine antibody function. Forfurther details, see Jones et al., Nature, 1986, 321:522-525; Riechmannet al., Nature, 1988, 332:323-329; and Presta, Curr. Op. Struct. Biol.,1992, 2:593-596, each of which is incorporated by reference in itsentirety.

A “human antibody” is one which possesses an amino acid sequencecorresponding to that of an antibody produced by a human or a humancell, or derived from a non-human source that utilizes a human antibodyrepertoire or human antibody-encoding sequences (e.g., obtained fromhuman sources or designed de novo). Human antibodies specificallyexclude humanized antibodies.

An “isolated antibody” is one that has been separated and/or recoveredfrom a component of its natural environment. Components of the naturalenvironment may include enzymes, hormones, and other proteinaceous ornonproteinaceous materials. In some embodiments, an isolated antibody ispurified to a degree sufficient to obtain at least 15 residues ofN-terminal or internal amino acid sequence, for example by use of aspinning cup sequenator. In some embodiments, an isolated antibody ispurified to homogeneity by gel electrophoresis (e.g., SDS-PAGE) underreducing or nonreducing conditions, with detection by Coomassie blue orsilver stain. An isolated antibody includes an antibody in situ withinrecombinant cells, since at least one component of the antibody'snatural environment is not present. In some aspects, an isolatedantibody is prepared by at least one purification step.

In some embodiments, an isolated antibody is purified to at least 80%,85%, 90%, 95%, or 99% by weight. In some embodiments, an isolatedantibody is provided as a solution comprising at least 85%, 90%, 95%,98%, 99% to 100% by weight of an antibody, the remainder of the weightcomprising the weight of other solutes dissolved in the solvent.

“Affinity” refers to the strength of the sum total of non-covalentinteractions between a single binding site of a molecule (e.g., anantibody) and its binding partner (e.g., an antigen). Unless indicatedotherwise, as used herein, “binding affinity” refers to intrinsicbinding affinity, which reflects a 1:1 interaction between members of abinding pair (e.g., antibody and antigen). The affinity of a molecule Xfor its partner Y can generally be represented by the dissociationconstant (K_(D)). Affinity can be measured by common methods known inthe art, including those described herein. Affinity can be determined,for example, using surface plasmon resonance (SPR) technology, such as aBiacore® instrument.

With regard to the binding of an antibody to a target molecule, theterms “specific binding,” “specifically binds to,” “specific for,”“selectively binds,” and “selective for” a particular antigen (e.g., apolypeptide target) or an epitope on a particular antigen mean bindingthat is measurably different from a non-specific or non-selectiveinteraction. Specific binding can be measured, for example, bydetermining binding of a molecule compared to binding of a controlmolecule. Specific binding can also be determined by competition with acontrol molecule that is similar to the target, such as an excess ofnon-labeled target. In that case, specific binding is indicated if thebinding of the labeled target to a probe is competitively inhibited bythe excess non-labeled target.

The term “k_(d)” (sec⁻¹), as used herein, refers to the dissociationrate constant of a particular antibody-antigen interaction. This valueis also referred to as the k_(off) value.

The term “k_(a)” (M⁻¹×sec⁻¹), as used herein, refers to the associationrate constant of a particular antibody-antigen interaction. This valueis also referred to as the k_(on) value.

The term “K_(D)” (M), as used herein, refers to the dissociationequilibrium constant of a particular antibody-antigen interaction.K_(D)=k_(d)/k_(a).

The term “K_(A)” (M⁻¹), as used herein, refers to the associationequilibrium constant of a particular antibody-antigen interaction.K_(A)=k_(a)/k_(d).

An “affinity matured” antibody is one with one or more alterations inone or more CDRs or FRs that result in an improvement in the affinity ofthe antibody for its antigen, compared to a parent antibody which doesnot possess the alteration(s). In one embodiment, an affinity maturedantibody has nanomolar or picomolar affinity for the target antigen.Affinity matured antibodies may be produced using a variety of methodsknown in the art. For example, Marks et al. (Bio/Technology, 1992,10:779-783, incorporated by reference in its entirety) describesaffinity maturation by V_(H) and V_(L) domain shuffling. Randommutagenesis of CDR and/or framework residues is described by, forexample, Barbas et al. (Proc. Nat. Acad. Sci. USA., 1994, 91:3809-3813);Schier et al., Gene, 1995, 169:147-155; Yelton et al., J. Immunol.,1995, 155:1994-2004; Jackson et al., J. Immunol., 1995, 154:3310-33199;and Hawkins et al, J. Mol. Biol., 1992, 226:889-896, each of which isincorporated by reference in its entirety.

When used herein in the context of two or more antibodies, the term“competes with” or “cross-competes with” indicates that the two or moreantibodies compete for binding to an antigen (e.g., PD-1). In oneexemplary assay, PD-1 is coated on a plate and allowed to bind a firstantibody, after which a second, labeled antibody is added. If thepresence of the first antibody reduces binding of the second antibody,then the antibodies compete. The term “competes with” also includescombinations of antibodies where one antibody reduces binding of anotherantibody, but where no competition is observed when the antibodies areadded in the reverse order. However, in some embodiments, the first andsecond antibodies inhibit binding of each other, regardless of the orderin which they are added. In some embodiments, one antibody reducesbinding of another antibody to its antigen by at least 50%, at least60%, at least 70%, at least 80%, or at least 90%.

The term “epitope” means a portion of an antigen capable of specificbinding to an antibody. Epitopes frequently consist ofsurface-accessible amino acid residues and/or sugar side chains and mayhave specific three dimensional structural characteristics, as well asspecific charge characteristics. Conformational and non-conformationalepitopes are distinguished in that the binding to the former but not thelatter is lost in the presence of denaturing solvents. An epitope maycomprise amino acid residues that are directly involved in the binding,and other amino acid residues, which are not directly involved in thebinding. The epitope to which an antibody binds can be determined usingknown techniques for epitope determination such as, for example, testingfor antibody binding to PD-1 variants with different point-mutations.

Percent “identity” between a polypeptide sequence and a referencesequence, is defined as the percentage of amino acid residues in thepolypeptide sequence that are identical to the amino acid residues inthe reference sequence, after aligning the sequences and introducinggaps, if necessary, to achieve the maximum percent sequence identity.Alignment for purposes of determining percent amino acid sequenceidentity can be achieved in various ways that are within the skill inthe art, for instance, using publicly available computer software suchas BLAST, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, or CLUSTAL OMEGAsoftware. Those skilled in the art can determine appropriate parametersfor aligning sequences, including any algorithms needed to achievemaximal alignment over the full length of the sequences being compared.

A “conservative substitution” or a “conservative amino acidsubstitution,” refers to the substitution of one or more amino acidswith one or more chemically or functionally similar amino acids.Conservative substitution tables providing similar amino acids are wellknown in the art. Polypeptide sequences having such substitutions areknown as “conservatively modified variants.” Such conservativelymodified variants are in addition to and do not exclude polymorphicvariants, interspecies homologs, and alleles. By way of example, thefollowing groups of amino acids are considered conservativesubstitutions for one another.

Acidic Residues D and E Basic Residues K, R, and H Hydrophilic UnchargedResidues S, T, N, and Q Aliphatic Uncharged Residues G, A, V, L, and INon-polar Uncharged Residues C, M, and P Aromatic Residues F, Y, and WAlcohol Group-Containing Residues S and T Aliphatic Residues I, L, V,and M Cycloalkenyl-associated Residues F, H, W, and Y HydrophobicResidues A, C, F, G, H, I, L, M, R, T, V, W, and Y Negatively ChargedResidues D and E Polar Residues C, D, E, H, K, N, Q, R, S, and TPositively Charged Residues H, K, and R Small Residues A, C, D, G, N, P,S, T, and V Very Small Residues A, G, and S Residues Involved in A, C,D, E, G, H, K, Turn Formation N, Q, R, S, P, and T Flexible Residues Q,T, K, S, G, P, D, E, and R Group 1 A, S, and T Group 2 D and E Group 3 Nand Q Group 4 R and K Group 5 I, L, and M Group 6 F, Y, and W Group A Aand G Group B D and E Group C N and Q Group D R, K, and H Group E I, L,M, V Group F F, Y, and W Group G S and T Group H C and MAdditional conservative substitutions may be found, for example, inCreighton, Proteins: Structures and Molecular Properties 2nd ed. (1993)W. H. Freeman & Co., New York, N.Y. An antibody generated by making oneor more conservative substitutions of amino acid residues in a parentantibody is referred to as a “conservatively modified variant.”

The term “amino acid” refers to the twenty common naturally occurringamino acids. Naturally occurring amino acids include alanine (Ala; A),arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine(Cys; C); glutamic acid (Glu; E), glutamine (Gln; Q), Glycine (Gly; G);histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys;K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P),serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr;Y), and valine (Val; V).

“Treating” or “treatment” of any disease or disorder refers, in certainembodiments, to ameliorating a disease or disorder that exists in asubject. In another embodiment, “treating” or “treatment” includesameliorating at least one physical parameter, which may be indiscernibleby the subject. In yet another embodiment, “treating” or “treatment”includes modulating the disease or disorder, either physically (e.g.,stabilization of a discernible symptom) or physiologically (e.g.,stabilization of a physical parameter) or both. In yet anotherembodiment, “treating” or “treatment” includes delaying or preventingthe onset of the disease or disorder.

As used herein, the term “therapeutically effective amount” or“effective amount” refers to an amount of an antibody or compositionthat when administered to a subject is effective to treat a disease ordisorder.

As used herein, the term “subject” means a mammalian subject. Exemplarysubjects include, but are not limited to humans, monkeys, dogs, cats,mice, rats, cows, horses, camels, avians, goats and sheep. In certainembodiments, the subject is a human. In some embodiments, the subjecthas cancer, an autoimmune disease or condition, and/or an infection thatcan be treated with an antibody provided herein. In some embodiments,the subject is a human that is suspected to have cancer, an autoimmunedisease or condition, and/or an infection.

2. Antibodies

Provided herein are antibodies that selectively bind human PD-1. In someaspects, the antibody selectively binds to the extracellular domain ofhuman PD-1. In some aspects, the antibody selectively binds to one ormore of full-length human PD-1, PD-1Δex2, PD-1Δex3, PD-1Δex2,3, andPD-1Δex2,3,4. See Nielsen et al., Cellular Immunology, 2005,235:109-116, incorporated by reference in its entirety.

In some embodiments, the antibody binds to homologs of human PD-1. Insome aspects, the antibody binds to a homolog of human PD-1 from aspecies selected from monkeys, mice, dogs, cats, rats, cows, horses,goats and sheep. In some aspects, the homolog is a cynomolgus monkeyhomolog. In some aspects, the homolog is a murine homolog.

In some embodiments, the antibody has one or more CDRs having particularlengths, in terms of the number of amino acid residues. In someembodiments, the Chothia CDR-H1 of the antibody is 6, 7, 8, or 9residues in length. In some embodiments, the Kabat CDR-H1 of theantibody is 4, 5, 6, or 7 residues in length. In some embodiments, theChothia CDR-H2 of the antibody is 5, 6, or 7 residues in length. In someembodiments, the Kabat CDR-H2 of the antibody is 15, 16, 17, or 18residues in length. In some embodiments, the Kabat/Chothia CDR-H3 of theantibody is 5, 6, 7, 8, 9, 10, 11, or 12 residues in length.

In some aspects, the Kabat/Chothia CDR-L1 of the antibody is 9, 10, 11,12, 13, 14, 15, or 16 residues in length. In some aspects, theKabat/Chothia CDR-L2 of the antibody is 6, 7, or 8 residues in length.In some aspects, the Kabat/Chothia CDR-L3 of the antibody is 8, 9, 10,11, or 12 residues in length.

In some embodiments, the antibody comprises a light chain. In someaspects, the light chain is a kappa light chain. In some aspects, thelight chain is a lambda light chain.

In some embodiments, the antibody comprises a heavy chain. In someaspects, the heavy chain is an IgA. In some aspects, the heavy chain isan IgD. In some aspects, the heavy chain is an IgE. In some aspects, theheavy chain is an IgG. In some aspects, the heavy chain is an IgM. Insome aspects, the heavy chain is an IgG1. In some aspects, the heavychain is an IgG2. In some aspects, the heavy chain is an IgG3. In someaspects, the heavy chain is an IgG4. In some aspects, the heavy chain isan IgA1. In some aspects, the heavy chain is an IgA2.

In some embodiments, the antibody is an antibody fragment. In someaspects, the antibody fragment is an Fv fragment. In some aspects, theantibody fragment is a Fab fragment. In some aspects, the antibodyfragment is a F(ab′)₂ fragment. In some aspects, the antibody fragmentis a Fab′ fragment. In some aspects, the antibody fragment is an scFv(sFv) fragment. In some aspects, the antibody fragment is an scFv-Fcfragment.

In some embodiments, the antibody is a monoclonal antibody. In someembodiments, the antibody is a polyclonal antibody.

In some embodiments, the antibody is a chimeric antibody. In someembodiments, the antibody is a humanized antibody. In some embodiments,the antibody is a human antibody.

In some embodiments, the antibody is an affinity matured antibody. Insome aspects, the antibody is an affinity matured antibody derived froman illustrative sequence provided in this disclosure.

In some embodiments, the antibody inhibits the binding of PD-1 to itsligands. In some aspects, the antibody inhibits the binding of PD-1 toPD-L1. In some aspects, the antibody inhibits the binding of PD-1 toPD-L2. In some aspects, the antibody inhibits the binding of PD-1 toPD-L1 and PD-L2.

The antibodies provided herein may be useful for the treatment of avariety of diseases and conditions, including cancers, autoimmunediseases, and infections.

2.1. CDR-H3 Sequences

In some embodiments, the antibody comprises a CDR-H3 sequencecomprising, consisting of, or consisting essentially of a sequenceselected from SEQ ID NOs: 113-131 and 309-315. In some aspects, theantibody comprises a CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 113. In some aspects, the antibodycomprises a CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 114. In some aspects, the antibody comprises aCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 115. In some aspects, the antibody comprises a CDR-H3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 116. In some aspects, the antibody comprises a CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 117.In some aspects, the antibody comprises a CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 118. In someaspects, the antibody comprises a CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 119. In some aspects, theantibody comprises a CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 120. In some aspects, the antibodycomprises a CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 121. In some aspects, the antibody comprises aCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 122. In some aspects, the antibody comprises a CDR-H3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 123. In some aspects, the antibody comprises a CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 124.In some aspects, the antibody comprises a CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 125. In someaspects, the antibody comprises a CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 126. In some aspects, theantibody comprises a CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 127. In some aspects, the antibodycomprises a CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 128. In some aspects, the antibody comprises aCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 129. In some aspects, the antibody comprises a CDR-H3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 130. In some aspects, the antibody comprises a CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 131.In some aspects, the antibody comprises a CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 309. In someaspects, the antibody comprises a CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 310. In some aspects, theantibody comprises a CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 311. In some aspects, the antibodycomprises a CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 312. In some aspects, the antibody comprises aCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 313. In some aspects, the antibody comprises a CDR-H3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 314. In some aspects, the antibody comprises a CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 315.

In some aspects, the CDR-H3 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-H3 sequence provided inthis disclosure. In some aspects, the CDR-H3 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H3sequences provided in this disclosure. In some aspects, the CDR-H3sequence comprises, consists of, or consists essentially of any of theillustrative CDR-H3 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

In some aspects, the CDR-H3 sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 132-136. Insome aspects, the CDR-H3 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 132. In some aspects, the CDR-H3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 133. In some aspects, the CDR-H3 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 134. In some aspects, theCDR-H3 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 135. In some aspects, the CDR-H3 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 136.

2.2. V_(H) Sequences Comprising Illustrative CDRs

In some embodiments, the antibody comprises a V_(H) sequence comprisingone or more CDR-H sequences comprising, consisting of, or consistingessentially of one or more illustrative CDR-H sequences provided in thisdisclosure, and variants thereof.

2.2.1. V_(H) Sequences Comprising Illustrative Kabat CDRs

In some embodiments, the antibody comprises a V_(H) sequence comprisingone or more Kabat CDR-H sequences comprising, consisting of, orconsisting essentially of one or more illustrative Kabat CDR-H sequencesprovided in this disclosure, and variants thereof.

2.2.1.1. Kabat CDR-H3

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H3 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 113-131 and 309-315.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 113. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 114. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 115.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 116. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 117. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 118.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 119. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 120. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 121.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 122. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 123. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 124.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 125. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 126. In some aspects, theantibody comprises a VII sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 127.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 128. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 129. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 130.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 131. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 309. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 310.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 311. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 312. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 313.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 314. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 315.

2.2.1.2. Kabat CDR-H2

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H2 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 84-102 or 331. Insome aspects, the antibody comprises a VII sequence comprising a KabatCDR-H2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 84. In some aspects, the antibody comprises a V_(H) sequencecomprising a Kabat CDR-H2 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 85. In some aspects, the antibodycomprises a V_(H) sequence comprising a Kabat CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 86.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 87. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H2 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 88. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 89.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 90. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H2 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 91. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 92.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 93. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H2 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 94. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 95.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 96. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H2 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 97. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 98.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 99. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H2 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 100. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 101.In some aspects, the antibody comprises a VII sequence comprising aKabat CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 102. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H2 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 331.

2.2.1.3. Kabat CDR-H1

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 31-49. In someaspects, the antibody comprises a V_(H) sequence comprising a KabatCDR-H1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 31. In some aspects, the antibody comprises a V_(H) sequencecomprising a Kabat CDR-H1 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 32. In some aspects, the antibodycomprises a V_(H) sequence comprising a Kabat CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 33.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 34 In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H1 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 35. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 36.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 37. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H1 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 38. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 39.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 40. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H1 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 41. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 42.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 43. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H1 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 44. In some aspects, theantibody comprises a V_(H) sequence comprising a Kabat CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 45.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 46. In some aspects, the antibody comprises aV_(H) sequence comprising a Kabat CDR-H1 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 47. In some aspects, theantibody comprises a VII sequence comprising a Kabat CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 48.In some aspects, the antibody comprises a V_(H) sequence comprising aKabat CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 49.

2.2.1.4. Kabat CDR-H3+Kabat CDR-H2

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H3 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 113-131 and 309-315,and a Kabat CDR-H2 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 84-102 or 331. Insome aspects, the Kabat CDR-H3 sequence and the Kabat CDR-H2 sequenceare both from a single illustrative V_(H) sequence provided in thisdisclosure. For example, in some aspects, the Kabat CDR-H3 and KabatCDR-H2 are both from a single illustrative V_(H) sequence selected fromSEQ ID NOs: 246-264 and 316-322.

2.2.1.5. Kabat CDR-H3+Kabat CDR-H1

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H3 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 113-131 and 309-315,and a Kabat CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 31-49. In someaspects, the Kabat CDR-H3 sequence and the Kabat CDR-H1 sequence areboth from a single illustrative V_(H) sequence provided in thisdisclosure. For example, in some aspects, the Kabat CDR-H3 and KabatCDR-H1 are both from a single illustrative VII sequence selected fromSEQ ID NOs: 246-264 and 316-322.

2.2.1.6. Kabat CDR-H1+Kabat CDR-H2

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 31-49 and a KabatCDR-H2 sequence comprising, consisting of, or consisting essentially ofa sequence selected from SEQ ID NOs: 84-102 or 331. In some aspects, theKabat CDR-H1 sequence and the Kabat CDR-H2 sequence are both from asingle illustrative V_(H) sequence provided in this disclosure. Forexample, in some aspects, the Kabat CDR-H1 and Kabat CDR-H2 are bothfrom a single illustrative V_(H) sequence selected from SEQ ID NOs:246-264. 2.2.1.7. Kabat CDR-H1+Kabat CDR-H2+Kabat CDR-H3

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Kabat CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 31-49, a KabatCDR-H2 sequence comprising, consisting of, or consisting essentially ofa sequence selected from SEQ ID NOs: 84-102 or 331, and a Kabat CDR-H3sequence comprising, consisting of, or consisting essentially of asequence selected from SEQ ID NOs: 113-131 and 309-315. In some aspects,the Kabat CDR-H1 sequence, Kabat CDR-H2 sequence, and Kabat CDR-H3sequence are all from a single illustrative VII sequence provided inthis disclosure. For example, in some aspects, the Kabat CDR-H1, KabatCDR-H2, and Kabat CDR-H3 are all from a single illustrative V_(H)sequence selected from SEQ ID NOs: 246-264 and 316-322.

2.2.1.8. Variants of V_(H) Sequences Comprising Illustrative Kabat CDRs

In some embodiments, the V_(H) sequences provided herein comprise avariant of an illustrative Kabat CDR-H3, CDR-H2, and/or CDR-H1 sequenceprovided in this disclosure.

In some aspects, the Kabat CDR-H3 sequence comprises, consists of, orconsists essentially of a variant of an illustrative Kabat CDR-H3sequence provided in this disclosure. In some aspects, the Kabat CDR-H3sequence comprises, consists of, or consists essentially of a sequencehaving at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of theillustrative Kabat CDR-H3 sequences provided in this disclosure. In someaspects, the Kabat CDR-H3 sequence comprises, consists of, or consistsessentially of any of the illustrative Kabat CDR-H3 sequences providedin this disclosure, with 1, 2, or 3 amino acid substitutions. In someaspects, the amino acid substitutions are conservative amino acidsubstitutions.

In some aspects, the Kabat CDR-H2 sequence comprises, consists of, orconsists essentially of a variant of an illustrative Kabat CDR-H2sequence provided in this disclosure. In some aspects, the Kabat CDR-H2sequence comprises, consists of, or consists essentially of a sequencehaving at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of theillustrative Kabat CDR-H2 sequences provided in this disclosure. In someaspects, the Kabat CDR-H2 sequence comprises, consists of, or consistsessentially of any of the illustrative Kabat CDR-H2 sequences providedin this disclosure, with 1, 2, or 3 amino acid substitutions. In someaspects, the amino acid substitutions are conservative amino acidsubstitutions.

In some aspects, the Kabat CDR-H1 sequence comprises, consists of, orconsists essentially of a variant of an illustrative Kabat CDR-H1sequence provided in this disclosure. In some aspects, the Kabat CDR-H1sequence comprises, consists of, or consists essentially of a sequencehaving at least 70%, 75%, 80%, 85%, 90%, or 95% identity with any of theillustrative Kabat CDR-H1 sequences provided in this disclosure. In someaspects, the Kabat CDR-H1 sequence comprises, consists of, or consistsessentially of any of the illustrative Kabat CDR-H1 sequences providedin this disclosure, with 1, 2, or 3 amino acid substitutions. In someaspects, the amino acid substitutions are conservative amino acidsubstitutions.

2.2.1.9. Excluded V_(H) Sequences Comprising Kabat CDRs

In some embodiments, the V_(H) sequences provided herein do not comprisecertain Kabat CDR-H3, CDR-H2, and/or CDR-H1 sequences.

In some aspects, the Kabat CDR-H3 sequence does not comprise, consistof, or consist essentially of a sequence selected from SEQ ID NOs:108-112 or 132-136. In some aspects, the Kabat CDR-H3 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 108. In someaspects, the Kabat CDR-H3 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 109. In some aspects, the Kabat CDR-H3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 110. In some aspects, the Kabat CDR-H3 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 111. In some aspects,the Kabat CDR-H3 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 112. In some aspects, the Kabat CDR-H3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 132. In some aspects, the Kabat CDR-H3 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 133. In some aspects,the Kabat CDR-H3 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 134. In some aspects, the Kabat CDR-H3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 135. In some aspects, the Kabat CDR-H3 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 136.

In some aspects, the Kabat CDR-H2 sequence does not comprise, consistof, or consist essentially of a sequence selected from SEQ ID NOs: 55-59or 103-107. In some aspects, the Kabat CDR-H2 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 55. In someaspects, the Kabat CDR-H2 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 56. In some aspects, the Kabat CDR-H2sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 57. In some aspects, the Kabat CDR-H2 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 58. In some aspects,the Kabat CDR-H2 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 59. In some aspects, the Kabat CDR-H2 sequencedoes not comprise, consist of, or consist essentially of SEQ ID NO: 103.In some aspects, the Kabat CDR-H2 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 104. In some aspects, the KabatCDR-H2 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 105. In some aspects, the Kabat CDR-H2 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 106. In someaspects, the Kabat CDR-H2 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 107.

In some aspects, the Kabat CDR-H1 sequence does not comprise, consistof, or consist essentially of a sequence selected from SEQ ID NOs: 2-6or 50-54. In some aspects, the Kabat CDR-H1 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 2. In some aspects, theKabat CDR-H1 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 3. In some aspects, the Kabat CDR-H1 sequencedoes not comprise, consist of, or consist essentially of SEQ ID NO: 4.In some aspects, the Kabat CDR-H1 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 5. In some aspects, the KabatCDR-H1 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 6. In some aspects, the Kabat CDR-H1 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 50. In someaspects, the Kabat CDR-H1 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 51. In some aspects, the Kabat CDR-H1sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 52. In some aspects, the Kabat CDR-H1 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 53. In some aspects,the Kabat CDR-H1 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 54.

2.2.2. V_(H) Sequences Comprising Illustrative Chothia CDRs

In some embodiments, the antibody comprises a V_(H) sequence comprisingone or more Chothia CDR-H sequences comprising, consisting of, orconsisting essentially of one or more illustrative Chothia CDR-Hsequences provided in this disclosure, and variants thereof.

2.2.2.1. Chothia CDR-H3

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H3 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 113-131 and 309-315.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 113. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 114. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 115. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 116. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 117.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 118. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 119. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 120. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 121. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 122.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 123. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 124. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 125. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 126. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 127.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 128. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 129. In someaspects, the antibody comprises a VII sequence comprising a ChothiaCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 130. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 131. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 309.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 310. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 311. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 312. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 313. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 314.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 315.

2.2.2.2. Chothia CDR-H2

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H2 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 60-78. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 60. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H2 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 61. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 62.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 63. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H2 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 64. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 65. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H2 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 66. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 67.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 68. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H2 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 69. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 70. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H2 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 71. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 72.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 73. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H2 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 74. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 75. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H2 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 76. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 77.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 78.

2.2.2.3. Chothia CDR-H1

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 7-25. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 7. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H1 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 8. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 9. Insome aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 10 In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H1 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 11. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 12. In some aspects, the antibody comprises a VII sequencecomprising a Chothia CDR-H1 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 13. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 14.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 15. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H1 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 16. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 17. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H1 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 18. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 19.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 20. In some aspects, the antibody comprises aV_(H) sequence comprising a Chothia CDR-H1 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 21. In someaspects, the antibody comprises a V_(H) sequence comprising a ChothiaCDR-H1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 22. In some aspects, the antibody comprises a V_(H) sequencecomprising a Chothia CDR-H1 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 23. In some aspects, the antibodycomprises a V_(H) sequence comprising a Chothia CDR-H1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 24.In some aspects, the antibody comprises a V_(H) sequence comprising aChothia CDR-H1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 25.

2.2.2.4. Chothia CDR-H3+Chothia CDR-H2

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H3 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 113-131 and 309-315,and a Chothia CDR-H2 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 60-78. In someaspects, the Chothia CDR-H3 sequence and the Chothia CDR-H2 sequence areboth from a single illustrative V_(H) sequence provided in thisdisclosure. For example, in some aspects, the Chothia CDR-H3 and ChothiaCDR-H2 are both from a single illustrative V_(H) sequence selected fromSEQ ID NOs: 246-264 and 316-322.

2.2.2.5. Chothia CDR-H3+Chothia CDR-H1

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H3 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 113-131 and 309-315,and a Chothia CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 7-25. In someaspects, the Chothia CDR-H3 sequence and the Chothia CDR-H1 sequence areboth from a single illustrative V_(H) sequence provided in thisdisclosure. For example, in some aspects, the Chothia CDR-H3 and ChothiaCDR-H1 are both from a single illustrative VII sequence selected fromSEQ ID NOs: 246-264 and 316-322.

2.2.2.6. Chothia CDR-H1+Chothia CDR-H2

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 7-25 and a ChothiaCDR-H2 sequence comprising, consisting of, or consisting essentially ofa sequence selected from SEQ ID NOs: 60-78. In some aspects, the ChothiaCDR-H1 sequence and the Chothia CDR-H2 sequence are both from a singleillustrative V_(H) sequence provided in this disclosure. For example, insome aspects, the Chothia CDR-H1 and Chothia CDR-H2 are both from asingle illustrative V_(H) sequence selected from SEQ ID NOs: 246-264.

2.2.2.7. Chothia CDR-H1+Chothia CDR-H2+Chothia CDR-H3

In some embodiments, the antibody comprises a V_(H) sequence comprisinga Chothia CDR-H1 sequence comprising, consisting of, or consistingessentially of a sequence selected from SEQ ID NOs: 7-25, a ChothiaCDR-H2 sequence comprising, consisting of, or consisting essentially ofa sequence selected from SEQ ID NOs: 60-78, and a Chothia CDR-H3sequence comprising, consisting of, or consisting essentially of asequence selected from SEQ ID NOs: 113-131 and 309-315. In some aspects,the Chothia CDR-H1 sequence, Chothia CDR-H2 sequence, and Chothia CDR-H3sequence are all from a single illustrative V_(H) sequence provided inthis disclosure. For example, in some aspects, the Chothia CDR-H1,Chothia CDR-H2, and Chothia CDR-H3 are all from a single illustrativeV_(H) sequence selected from SEQ ID NOs: 246-264 and 316-322.

2.2.2.8. Variants of V_(H) Sequences Comprising Illustrative ChothiaCDRs

In some embodiments, the V_(H) sequences provided herein comprise avariant of an illustrative Chothia CDR-H3, CDR-H2, and/or CDR-H1sequence provided in this disclosure.

In some aspects, the Chothia CDR-H3 sequence comprises, consists of, orconsists essentially of a variant of an illustrative Chothia CDR-H3sequence provided in this disclosure. In some aspects, the ChothiaCDR-H3 sequence comprises, consists of, or consists essentially of asequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity withany of the illustrative Chothia CDR-H3 sequences provided in thisdisclosure. In some aspects, the Chothia CDR-H3 sequence comprises,consists of, or consists essentially of any of the illustrative ChothiaCDR-H3 sequences provided in this disclosure, with 1, 2, or 3 amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

In some aspects, the Chothia CDR-H2 sequence comprises, consists of, orconsists essentially of a variant of an illustrative Chothia CDR-H2sequence provided in this disclosure. In some aspects, the ChothiaCDR-H2 sequence comprises, consists of, or consists essentially of asequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity withany of the illustrative Chothia CDR-H2 sequences provided in thisdisclosure. In some aspects, the Chothia CDR-H2 sequence comprises,consists of, or consists essentially of any of the illustrative ChothiaCDR-H2 sequences provided in this disclosure, with 1, 2, or 3 amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

In some aspects, the Chothia CDR-H1 sequence comprises, consists of, orconsists essentially of a variant of an illustrative Chothia CDR-H1sequence provided in this disclosure. In some aspects, the ChothiaCDR-H1 sequence comprises, consists of, or consists essentially of asequence having at least 70%, 75%, 80%, 85%, 90%, or 95% identity withany of the illustrative Chothia CDR-H1 sequences provided in thisdisclosure. In some aspects, the Chothia CDR-H1 sequence comprises,consists of, or consists essentially of any of the illustrative ChothiaCDR-H1 sequences provided in this disclosure, with 1, 2, or 3 amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

2.2.2.9. Excluded V_(H) Sequences Comprising Chothia CDRs

In some embodiments, the V_(H) sequences provided herein do not comprisecertain Chothia CDR-H3, CDR-H2, and/or CDR-H1 sequences.

In some aspects, the Chothia CDR-H3 sequence does not comprise, consistof, or consist essentially of a sequence selected from SEQ ID NOs:108-112 or 132-136. In some aspects, the Chothia CDR-H3 sequence doesnot comprise, consist of, or consist essentially of SEQ ID NO: 108. Insome aspects, the Chothia CDR-H3 sequence does not comprise, consist of,or consist essentially of SEQ ID NO: 109. In some aspects, the ChothiaCDR-H3 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 110. In some aspects, the Chothia CDR-H3 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 111. In someaspects, the Chothia CDR-H3 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 112. In some aspects, the ChothiaCDR-H3 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 132. In some aspects, the Chothia CDR-H3 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 133. In someaspects, the Chothia CDR-H3 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 134. In some aspects, the ChothiaCDR-H3 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 135. In some aspects, the Chothia CDR-H3 sequence does notcomprise, consist of, or consist essentially of SEQ ID NO: 136.

In some aspects, the Chothia CDR-H2 sequence does not comprise, consistof, or consist essentially of a sequence selected from SEQ ID NOs:79-83. In some aspects, the Chothia CDR-H2 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 79. In some aspects,the Chothia CDR-H2 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 80. In some aspects, the Chothia CDR-H2sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 81. In some aspects, the Chothia CDR-H2 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 82. In some aspects,the Chothia CDR-H2 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 83.

In some aspects, the Chothia CDR-H1 sequence does not comprise, consistof, or consist essentially of a sequence selected from SEQ ID NOs:26-30. In some aspects, the Chothia CDR-H1 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 26. In some aspects,the Chothia CDR-H1 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 27. In some aspects, the Chothia CDR-H1sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 28. In some aspects, the Chothia CDR-H1 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 29. In some aspects,the Chothia CDR-H1 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 30.

2.3. V_(H) Sequences

In some embodiments, the antibody comprises a V_(H) sequence comprising,consisting of, or consisting essentially of a sequence selected from SEQID NOs: 246-264 and 316-322. In some aspects, the antibody comprises aV_(H) sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 246. In some aspects, the antibody comprises a V_(H) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 247.In some aspects, the antibody comprises a V_(H) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 248. In someaspects, the antibody comprises a V_(H) sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 249. In some aspects, theantibody comprises a V_(H) sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 250. In some aspects, the antibodycomprises a V_(H) sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 251. In some aspects, the antibody comprises aV_(H) sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 252. In some aspects, the antibody comprises a V_(H) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 253.In some aspects, the antibody comprises a V_(H) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 254. In someaspects, the antibody comprises a V_(H) sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 255 (with or without aserine prepended to the sequence). In some aspects, the antibodycomprises a V_(H) sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 256. In some aspects, the antibody comprises aV_(H) sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 257. In some aspects, the antibody comprises a V_(H) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 258.In some aspects, the antibody comprises a V_(H) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 259. In someaspects, the antibody comprises a V_(H) sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 260. In some aspects, theantibody comprises a V_(H) sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 261. In some aspects, the antibodycomprises a V_(H) sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 262. In some aspects, the antibody comprises aV_(H) sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 263. In some aspects, the antibody comprises a V_(H) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 264.In some aspects, the antibody comprises a V_(H) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 316. In someaspects, the antibody comprises a V_(H) sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 317. In some aspects, theantibody comprises a V_(H) sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 318. In some aspects, the antibodycomprises a V_(H) sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 319. In some aspects, the antibody comprises aV_(H) sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 320. In some aspects, the antibody comprises a V_(H) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 321.In some aspects, the antibody comprises a V_(H) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 322.

2.3.1. Variants of V_(H) Sequences

In some embodiments, the V_(H) sequences provided herein comprise,consist of, or consist essentially of a variant of an illustrative V_(H)sequence provided in this disclosure.

In some aspects, the V_(H) sequence comprises, consists of, or consistsessentially of a variant of an illustrative V_(H) sequence provided inthis disclosure. In some aspects, the V_(H) sequence comprises, consistsof, or consists essentially of a sequence having at least 85%, 90%, 95%,96%, 97%, 98%, 99%, or 99.5% identity with any of the illustrative V_(H)sequences provided in this disclosure.

In some embodiments, the V_(H) sequence comprises, consists of, orconsists essentially of any of the illustrative V_(H) sequences providedin this disclosure, 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer,16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 orfewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 orfewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

2.3.2. Excluded V_(H) Sequences

In some embodiments, the V_(H) sequences provided herein do not comprisecertain V_(H) sequences.

In some aspects, the V_(H) sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 265-269. Insome aspects, the V_(H) sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 265. In some aspects, the V_(H)sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 266. In some aspects, the V_(H) sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 267. In some aspects, the V_(H)sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 268. In some aspects, the V_(H) sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 269.

2.4. CDR-L3 Sequences

In some embodiments, the antibody comprises a CDR-L3 sequencecomprising, consisting of, or consisting essentially of a sequenceselected from SEQ ID NOs: 200-218. In some aspects, the antibodycomprises a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 200. In some aspects, the antibody comprises aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 201. In some aspects, the antibody comprises a CDR-L3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 202. In some aspects, the antibody comprises a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 203.In some aspects, the antibody comprises a CDR-L3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 204. In someaspects, the antibody comprises a CDR-L3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 205. In some aspects, theantibody comprises a CDR-L3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 206. In some aspects, the antibodycomprises a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 207. In some aspects, the antibody comprises aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 208. In some aspects, the antibody comprises a CDR-L3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 209. In some aspects, the antibody comprises a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 210.In some aspects, the antibody comprises a CDR-L3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 211. In someaspects, the antibody comprises a CDR-L3 sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 212. In some aspects, theantibody comprises a CDR-L3 sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 213. In some aspects, the antibodycomprises a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 214. In some aspects, the antibody comprises aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 215. In some aspects, the antibody comprises a CDR-L3sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 216. In some aspects, the antibody comprises a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 217.In some aspects, the antibody comprises a CDR-L3 sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 218.

In some aspects, the CDR-L3 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-L3 sequence provided inthis disclosure. In some aspects, the CDR-L3 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3sequences provided in this disclosure. In some aspects, the CDR-L3sequence comprises, consists of, or consists essentially of any of theillustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

In some aspects, the CDR-L3 sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 195-199 or219-223. In some aspects the CDR-L3 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 195. In some aspects the CDR-L3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 196. In some aspects the CDR-L3 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 197. In some aspects the CDR-L3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 198. In some aspects the CDR-L3 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 199. In some aspects the CDR-L3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 219. In some aspects the CDR-L3 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 220. In some aspects the CDR-L3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 221. In some aspects the CDR-L3 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 222. In some aspects the CDR-L3sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 223.

2.5. V_(L) Sequences Comprising Illustrative CDRs

In some embodiments, the antibody comprises a V_(L) sequence comprisingone or more CDR-L sequences comprising, consisting of, or consistingessentially of one or more illustrative CDR-L sequences provided in thisdisclosure, and variants thereof.

2.5.1. CDR-L3

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L3 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 200-218. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 200.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 201. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 202. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L3 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 203. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 204.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 205. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 206. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L3 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 207. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 208.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 209. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 210. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L3 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 211. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 212.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 213. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 214. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L3 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 215. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L3 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 216.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L3 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 217. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L3 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 218.

2.5.2. CDR-L2

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L2 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 171-189. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 171.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 172. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 173. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L2 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 174. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 175.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 176. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 177. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L2 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 178. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 179.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 180. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 181. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L2 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 182. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 183.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 184. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 185. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L2 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 186. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L2 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 187.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L2 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 188. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L2 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 189.

2.5.3. CDR-L1

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L1 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 142-160. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 142.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 143. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 144. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L1 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 145. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 146.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 147. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 148. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L1 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 149. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 150.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 151. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 152. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L1 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 153. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 154.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 155. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 156. In some aspects, the antibody comprises aV_(L) sequence comprising a CDR-L1 sequence comprising, consisting of,or consisting essentially of SEQ ID NO: 157. In some aspects, theantibody comprises a V_(L) sequence comprising a CDR-L1 sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 158.In some aspects, the antibody comprises a V_(L) sequence comprising aCDR-L1 sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 159. In some aspects, the antibody comprises a V_(L) sequencecomprising a CDR-L1 sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 160.

2.5.4. CDR-L3+CDR-L2

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L3 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 200-218 and a CDR-L2 sequencecomprising, consisting of, or consisting essentially of a sequenceselected from SEQ ID NOs: 171-189. In some aspects, the CDR-L3 sequenceand the CDR-L2 sequence are both from a single illustrative V_(L)sequence provided in this disclosure. For example, in some aspects, theCDR-L3 and CDR-L2 are both from a single illustrative V_(L) sequenceselected from SEQ ID NOs: 270-288.

2.5.5. CDR-L3+CDR-L1

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L3 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 200-218 and a CDR-L1 sequencecomprising, consisting of, or consisting essentially of a sequenceselected from SEQ ID NOs: 142-160. In some aspects, the CDR-L3 sequenceand the CDR-L1 sequence are both from a single illustrative V_(L)sequence provided in this disclosure. For example, in some aspects, theCDR-L3 and CDR-L1 are both from a single illustrative V_(L) sequenceselected from SEQ ID NOs: 270-288.

2.5.6. CDR-L1+CDR-L2

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L1 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 142-160 and a CDR-L2 sequencecomprising, consisting of, or consisting essentially of a sequenceselected from SEQ ID NOs: 171-189. In some aspects, the CDR-L1 sequenceand the CDR-L2 sequence are both from a single illustrative V_(L)sequence provided in this disclosure. For example, in some aspects, theCDR-L1 and CDR-L2 are both from a single illustrative V_(L) sequenceselected from SEQ ID NOs: 270-288.

2.5.7. CDR-L1+CDR-L2+CDR-L3

In some embodiments, the antibody comprises a V_(L) sequence comprisinga CDR-L1 sequence comprising, consisting of, or consisting essentiallyof a sequence selected from SEQ ID NOs: 142-160, a CDR-L2 sequencecomprising, consisting of, or consisting essentially of a sequenceselected from SEQ ID NOs: 171-189, and a CDR-L3 sequence comprising,consisting of, or consisting essentially of a sequence selected from SEQID NOs: 200-218. In some aspects, the CDR-L1 sequence, CDR-L2 sequence,and CDR-L3 sequence are all from a single illustrative V_(L) sequenceprovided in this disclosure. For example, in some aspects, the CDR-L1,CDR-L2, and CDR-L3 are all from a single illustrative V_(L) sequenceselected from SEQ ID NOs: 270-288.

2.5.8. Variants of V_(L) Sequences Comprising Illustrative CDR-Ls

In some embodiments, the V_(L) sequences provided herein comprise avariant of an illustrative CDR-L3, CDR-L2, and/or CDR-L1 sequenceprovided in this disclosure.

In some aspects, the CDR-L3 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-L3 sequence provided inthis disclosure. In some aspects, the CDR-L3 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3sequences provided in this disclosure. In some aspects, the CDR-L3sequence comprises, consists of, or consists essentially of any of theillustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

In some aspects, the CDR-L2 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-L2 sequence provided inthis disclosure. In some aspects, the CDR-L2 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L2sequences provided in this disclosure. In some aspects, the CDR-L2sequence comprises, consists of, or consists essentially of any of theillustrative CDR-L2 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

In some aspects, the CDR-L1 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-L1 sequence provided inthis disclosure. In some aspects, the CDR-L1 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L1sequences provided in this disclosure. In some aspects, the CDR-L1sequence comprises, consists of, or consists essentially of any of theillustrative CDR-L1 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

2.5.9. Excluded V_(L) Sequences Comprising CDR-Ls

In some embodiments, the V_(L) sequences provided herein do not comprisecertain CDR-L3, CDR-L2, and/or CDR-L1 sequences.

In some aspects, the CDR-L3 sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 195-199 or219-223. In some aspects, the CDR-L3 sequence does not comprise, consistof, or consist essentially of SEQ ID NOs: 195. In some aspects, theCDR-L3 sequence does not comprise, consist of, or consist essentially ofSEQ ID NOs: 196. In some aspects, the CDR-L3 sequence does not comprise,consist of, or consist essentially of SEQ ID NOs: 197. In some aspects,the CDR-L3 sequence does not comprise, consist of, or consistessentially of SEQ ID NOs: 198. In some aspects, the CDR-L3 sequencedoes not comprise, consist of, or consist essentially of SEQ ID NOs:199. In some aspects, the CDR-L3 sequence does not comprise, consist of,or consist essentially of SEQ ID NOs: 219. In some aspects, the CDR-L3sequence does not comprise, consist of, or consist essentially of SEQ IDNOs: 220. In some aspects, the CDR-L3 sequence does not comprise,consist of, or consist essentially of SEQ ID NOs: 221. In some aspects,the CDR-L3 sequence does not comprise, consist of, or consistessentially of SEQ ID NOs: 222. In some aspects, the CDR-L3 sequencedoes not comprise, consist of, or consist essentially of SEQ ID NOs:223.

In some aspects, the CDR-L2 sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 166-170 or190-194. In some aspects, the CDR-L2 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 190. In some aspects, theCDR-L2 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 166. In some aspects, the CDR-L2 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 167. In some aspects,the CDR-L2 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 168. In some aspects, the CDR-L2 sequence doesnot comprise, consist of, or consist essentially of SEQ ID NO: 169. Insome aspects, the CDR-L2 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 170. In some aspects, the CDR-L2sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 191. In some aspects, the CDR-L2 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 192. In some aspects, theCDR-L2 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 193. In some aspects, the CDR-L2 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 194.

In some aspects, the CDR-L1 sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 137-141 or161-165. In some aspects, the CDR-L1 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 137. In some aspects, theCDR-L1 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 138. In some aspects, the CDR-L1 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 139. In some aspects,the CDR-L1 sequence does not comprise, consist of, or consistessentially of SEQ ID NO: 140. In some aspects, the CDR-L1 sequence doesnot comprise, consist of, or consist essentially of SEQ ID NO: 141. Insome aspects, the CDR-L1 sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 161. In some aspects, the CDR-L1sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 162. In some aspects, the CDR-L1 sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 163. In some aspects, theCDR-L1 sequence does not comprise, consist of, or consist essentially ofSEQ ID NO: 164. In some aspects, the CDR-L1 sequence does not comprise,consist of, or consist essentially of SEQ ID NO: 165.

2.6. V_(L) Sequences

In some embodiments, the antibody comprises a V_(L) sequence comprising,consisting of, or consisting essentially of a sequence selected from SEQID NOs: 270-288. In some aspects, the antibody comprises a V_(L)sequence comprising, consisting of, or consisting essentially of SEQ IDNO: 270. In some aspects, the antibody comprises a V_(L) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 281.In some aspects, the antibody comprises a V_(L) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 282. In someaspects, the antibody comprises a V_(L) sequence comprising, consistingof, or consisting essentially of SEQ ID NO: 283. In some aspects, theantibody comprises a V_(L) sequence comprising, consisting of, orconsisting essentially of SEQ ID NO: 284. In some aspects, the antibodycomprises a V_(L) sequence comprising, consisting of, or consistingessentially of SEQ ID NO: 285. In some aspects, the antibody comprises aV_(L) sequence comprising, consisting of, or consisting essentially ofSEQ ID NO: 286. In some aspects, the antibody comprises a V_(L) sequencecomprising, consisting of, or consisting essentially of SEQ ID NO: 287.In some aspects, the antibody comprises a V_(L) sequence comprising,consisting of, or consisting essentially of SEQ ID NO: 288.

2.6.1. Variants of V_(L) Sequences

In some embodiments, the V_(L) sequences provided herein comprise,consist of, or consist essentially of a variant of an illustrative V_(L)sequence provided in this disclosure.

In some aspects, the V_(L) sequence comprises, consists of, or consistsessentially of a variant of an illustrative V_(L) sequence provided inthis disclosure. In some aspects, the V_(L) sequence comprises, consistsof, or consists essentially of a sequence having at least 85%, 90%, 95%,96%, 97%, 98%, 99%, or 99.05% identity with any of the illustrativeV_(L) sequences provided in this disclosure.

In some embodiments, the V_(L) sequence comprises, consists of, orconsists essentially of any of the illustrative V_(L) sequences providedin this disclosure, 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer,16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 orfewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 orfewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

2.6.2. Excluded V_(L) Sequences

In some embodiments, the V_(L) sequences provided herein do not comprisecertain V_(L) sequences.

In some aspects, the V_(L) sequence does not comprise, consist of, orconsist essentially of a sequence selected from SEQ ID NOs: 289-293. Insome aspects, the V_(L) sequence does not comprise, consist of, orconsist essentially of SEQ ID NO: 289. In some aspects, the V_(L)sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 290. In some aspects, the V_(L) sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 291. In some aspects, the V_(L)sequence does not comprise, consist of, or consist essentially of SEQ IDNO: 292. In some aspects, the V_(L) sequence does not comprise, consistof, or consist essentially of SEQ ID NO: 293.

2.7. Pairs

2.7.1. CDR-H3-CDR-L3 Pairs

In some embodiments, the antibody comprises a CDR-H3 sequence and aCDR-L3 sequence. In some aspects, the CDR-H3 sequence is part of a VIIand the CDR-L3 sequence is part of a V_(L).

In some aspects, the CDR-H3 sequence is a CDR-H3 sequence comprising,consisting of, or consisting essentially of SEQ ID NOs: 113-131 and309-315, and the CDR-L3 sequence is a CDR-L3 sequence comprising,consisting of, or consisting essentially of SEQ ID NOs: 200-218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 113, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 114, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 115, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 116, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 117, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 118, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 119, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 120, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 121, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 122, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 123, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 124, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 125, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 126, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 127, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 128, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 129, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 130, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 131, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 309, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 310, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 311, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 312, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 313, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 314, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

In some aspects, the CDR-H3 sequence is SEQ ID NO: 315, and the CDR-L3sequence is selected from SEQ ID NOs: 200-218. In some aspects, theCDR-L3 sequence is SEQ ID NO: 200. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 201. In some aspects, the CDR-L3 sequence is SEQ ID NO:202. In some aspects, the CDR-L3 sequence is SEQ ID NO: 203. In someaspects, the CDR-L3 sequence is SEQ ID NO: 204. In some aspects, theCDR-L3 sequence is SEQ ID NO: 205. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 206. In some aspects, the CDR-L3 sequence is SEQ ID NO:207. In some aspects, the CDR-L3 sequence is SEQ ID NO: 208. In someaspects, the CDR-L3 sequence is SEQ ID NO: 209. In some aspects, theCDR-L3 sequence is SEQ ID NO: 210. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 211. In some aspects, the CDR-L3 sequence is SEQ ID NO:212. In some aspects, the CDR-L3 sequence is SEQ ID NO: 213. In someaspects, the CDR-L3 sequence is SEQ ID NO: 214. In some aspects, theCDR-L3 sequence is SEQ ID NO: 215. In some aspects, the CDR-L3 sequenceis SEQ ID NO: 216. In some aspects, the CDR-L3 sequence is SEQ ID NO:217. In some aspects, the CDR-L3 sequence is SEQ ID NO: 218.

2.7.1.1. Variants of CDR-H3-CDR-L3 Pairs

In some embodiments, the CDR-H3-CDR-L3 pairs provided herein comprise avariant of an illustrative CDR-H3 and/or CDR-L1 sequence provided inthis disclosure.

In some aspects, the CDR-H3 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-H3 sequence provided inthis disclosure. In some aspects, the CDR-H3 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-H3sequences provided in this disclosure. In some aspects, the CDR-H3sequence comprises, consists of, or consists essentially of any of theillustrative CDR-H3 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

In some aspects, the CDR-L3 sequence comprises, consists of, or consistsessentially of a variant of an illustrative CDR-L3 sequence provided inthis disclosure. In some aspects, the CDR-L3 sequence comprises,consists of, or consists essentially of a sequence having at least 70%,75%, 80%, 85%, 90%, or 95% identity with any of the illustrative CDR-L3sequences provided in this disclosure. In some aspects, the CDR-L3sequence comprises, consists of, or consists essentially of any of theillustrative CDR-L3 sequences provided in this disclosure, with 1, 2, or3 amino acid substitutions. In some aspects, the amino acidsubstitutions are conservative amino acid substitutions.

2.7.1.2. Excluded CDR-H3-CDR-L3 Pairs

In some embodiments, the CDR-H3-CDR-L3 pairs provided herein do notcomprise certain CDR-H3-CDR-L3 pairs.

In some aspects, the CDR-H3 sequence is not selected from SEQ ID NOs:108-112 or 132-136, and the CDR-L3 sequence is not selected from SEQ IDNOs: 195-199 or 219-223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 108, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 109, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 110, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 111, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 112, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 132, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 133, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 134, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 135, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

In some aspects, the CDR-H3 sequence is not SEQ ID NO: 136, and theCDR-L3 sequence is not selected from SEQ ID NOs: 195-199 or 219-223. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 195. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 196. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 197. In some aspects, the CDR-L3sequence is not SEQ ID NO: 198. In some aspects, the CDR-L3 sequence isnot SEQ ID NO: 199. In some aspects, the CDR-L3 sequence is not SEQ IDNO: 219. In some aspects, the CDR-L3 sequence is not SEQ ID NO: 220. Insome aspects, the CDR-L3 sequence is not SEQ ID NO: 221. In someaspects, the CDR-L3 sequence is not SEQ ID NO: 222. In some aspects, theCDR-L3 sequence is not SEQ ID NO: 223.

2.7.2. V_(H)-V_(L) Pairs

In some embodiments, the antibody comprises a V_(H) sequence and a V_(L)sequence.

In some aspects, the V_(H) sequence is a V_(H) sequence comprising,consisting of, or consisting essentially of SEQ ID NOs: 246-264 and316-322, and the V_(L) sequence is a V_(L) sequence comprising,consisting of, or consisting essentially of SEQ ID NOs: 270-288.

In some aspects, the V_(H) sequence is SEQ ID NO: 246, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 247, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 248, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 249, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 250, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 251, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 252, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 253, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 254, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 255 (with or without aserine prepended to the sequence), and the V_(L) sequence is selectedfrom SEQ ID NOs: 270-288. In some aspects, the V_(L) sequence is SEQ IDNO: 270. In some aspects, the V_(L) sequence is SEQ ID NO: 271. In someaspects, the V_(L) sequence is SEQ ID NO: 272. In some aspects, theV_(L) sequence is SEQ ID NO: 273. In some aspects, the V_(L) sequence isSEQ ID NO: 274. In some aspects, the V_(L) sequence is SEQ ID NO: 275.In some aspects, the V_(L) sequence is SEQ ID NO: 276. In some aspects,the V_(L) sequence is SEQ ID NO: 277. In some aspects, the V_(L)sequence is SEQ ID NO: 278. In some aspects, the V_(L) sequence is SEQID NO: 279. In some aspects, the V_(L) sequence is SEQ ID NO: 280. Insome aspects, the V_(L) sequence is SEQ ID NO: 281. In some aspects, theV_(L) sequence is SEQ ID NO: 282. In some aspects, the V_(L) sequence isSEQ ID NO: 283. In some aspects, the V_(L) sequence is SEQ ID NO: 284.In some aspects, the V_(L) sequence is SEQ ID NO: 285. In some aspects,the V_(L) sequence is SEQ ID NO: 286. In some aspects, the V_(L)sequence is SEQ ID NO: 287. In some aspects, the V_(L) sequence is SEQID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 256, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 257, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 258, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 259, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 260, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 261, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 262, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 263, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 264, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 316, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 317, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 318, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 319, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 320, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 321, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

In some aspects, the V_(H) sequence is SEQ ID NO: 322, and the V_(L)sequence is selected from SEQ ID NOs: 270-288. In some aspects, theV_(L) sequence is SEQ ID NO: 270. In some aspects, the V_(L) sequence isSEQ ID NO: 271. In some aspects, the V_(L) sequence is SEQ ID NO: 272.In some aspects, the V_(L) sequence is SEQ ID NO: 273. In some aspects,the V_(L) sequence is SEQ ID NO: 274. In some aspects, the V_(L)sequence is SEQ ID NO: 275. In some aspects, the V_(L) sequence is SEQID NO: 276. In some aspects, the V_(L) sequence is SEQ ID NO: 277. Insome aspects, the V_(L) sequence is SEQ ID NO: 278. In some aspects, theV_(L) sequence is SEQ ID NO: 279. In some aspects, the V_(L) sequence isSEQ ID NO: 280. In some aspects, the V_(L) sequence is SEQ ID NO: 281.In some aspects, the V_(L) sequence is SEQ ID NO: 282. In some aspects,the V_(L) sequence is SEQ ID NO: 283. In some aspects, the V_(L)sequence is SEQ ID NO: 284. In some aspects, the V_(L) sequence is SEQID NO: 285. In some aspects, the V_(L) sequence is SEQ ID NO: 286. Insome aspects, the V_(L) sequence is SEQ ID NO: 287. In some aspects, theV_(L) sequence is SEQ ID NO: 288.

2.7.2.1. Variants of V_(H)-V_(L) Pairs

In some embodiments, the V_(H)-V_(L) pairs provided herein comprise avariant of an illustrative V_(H) and/or V_(L) sequence provided in thisdisclosure.

In some aspects, the V_(H) sequence comprises, consists of, or consistsessentially of a variant of an illustrative V_(H) sequence provided inthis disclosure. In some aspects, the V_(H) sequence comprises, consistsof, or consists essentially of a sequence having at least 85%, 90%, 95%,96%, 97%, 98%, 99%, or 99.1% identity with any of the illustrative V_(H)sequences provided in this disclosure.

In some embodiments, the V_(H) sequence comprises, consists of, orconsists essentially of any of the illustrative V_(H) sequences providedin this disclosure, 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer,16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 orfewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 orfewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

In some aspects, the V_(L) sequence comprises, consists of, or consistsessentially of a variant of an illustrative V_(L) sequence provided inthis disclosure. In some aspects, the V_(L) sequence comprises, consistsof, or consists essentially of a sequence having at least 85%, 90%, 95%,96%, 97%, 98%, 99%, or 99.05% identity with any of the illustrativeV_(L) sequences provided in this disclosure.

In some embodiments, the V_(L) sequence comprises, consists of, orconsists essentially of any of the illustrative V_(L) sequences providedin this disclosure, 20 or fewer, 19 or fewer, 18 or fewer, 17 or fewer,16 or fewer, 15 or fewer, 14 or fewer, 13 or fewer, 12 or fewer, 11 orfewer, 10 or fewer, 9 or fewer, 8 or fewer, 7 or fewer, 6 or fewer, 5 orfewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer amino acidsubstitutions. In some aspects, the amino acid substitutions areconservative amino acid substitutions.

2.7.2.2. Excluded V_(H)-V_(L) Pairs

In some embodiments, the V_(H)-V_(L) pairs provided herein do notcomprise certain V_(H)-V_(L) pairs.

In some aspects, the V_(H) sequence is not selected from SEQ ID NOs:265-269, and the V_(L) sequence is not selected from SEQ ID NOs:289-293.

In some aspects, the V_(H) sequence is not SEQ ID NO: 265, and the V_(L)sequence is not selected from SEQ ID NO: 289-293. In some aspects, theV_(L) sequence is not SEQ ID NO: 289. In some aspects, the V_(L)sequence is not SEQ ID NO: 290. In some aspects, the V_(L) sequence isnot SEQ ID NO: 291. In some aspects, the V_(L) sequence is not SEQ IDNO: 292. In some aspects, the V_(L) sequence is not SEQ ID NO: 293.

In some aspects, the V_(H) sequence is not SEQ ID NO: 266, and the V_(L)sequence is not selected from SEQ ID NO: 289-293. In some aspects, theV_(L) sequence is not SEQ ID NO: 289. In some aspects, the V_(L)sequence is not SEQ ID NO: 290. In some aspects, the V_(L) sequence isnot SEQ ID NO: 291. In some aspects, the V_(L) sequence is not SEQ IDNO: 292. In some aspects, the V_(L) sequence is not SEQ ID NO: 293.

In some aspects, the V_(H) sequence is not SEQ ID NO: 267, and the V_(L)sequence is not selected from SEQ ID NO: 289-293. In some aspects, theV_(L) sequence is not SEQ ID NO: 289. In some aspects, the V_(L)sequence is not SEQ ID NO: 290. In some aspects, the V_(L) sequence isnot SEQ ID NO: 291. In some aspects, the V_(L) sequence is not SEQ IDNO: 292. In some aspects, the V_(L) sequence is not SEQ ID NO: 293.

In some aspects, the V_(H) sequence is not SEQ ID NO: 268, and the V_(L)sequence is not selected from SEQ ID NO: 289-293. In some aspects, theV_(L) sequence is not SEQ ID NO: 289. In some aspects, the V_(L)sequence is not SEQ ID NO: 290. In some aspects, the V_(L) sequence isnot SEQ ID NO: 291. In some aspects, the V_(L) sequence is not SEQ IDNO: 292. In some aspects, the V_(L) sequence is not SEQ ID NO: 293.

In some aspects, the V_(H) sequence is not SEQ ID NO: 269, and the V_(L)sequence is not selected from SEQ ID NO: 289-293. In some aspects, theV_(L) sequence is not SEQ ID NO: 289. In some aspects, the V_(L)sequence is not SEQ ID NO: 290. In some aspects, the V_(L) sequence isnot SEQ ID NO: 291. In some aspects, the V_(L) sequence is not SEQ IDNO: 292. In some aspects, the V_(L) sequence is not SEQ ID NO: 293.

2.8. Consensus Sequences

In some embodiments, provided herein are anti-PD-1 antibodies comprisingone or more sequences defined by consensus sequences. Each consensussequence is based, at least in part, on one or more alignments of two ormore useful anti-PD-1 CDR sequences provided in this disclosure. Basedon such alignments, a person of skill in the art would recognize thatdifferent amino acid residues may useful in certain positions of theCDRs. Accordingly, each consensus sequence encompasses two or moreuseful anti-PD-1 CDR sequences.

2.8.1. CDR-H3 Consensus Sequences

In some embodiments, the antibody comprises a CDR-H3 sequence defined bythe consensus sequence D-α₂-α₃-Y-α₅-α₆-G-S-G-Y, where α₂ is A, V, or S;α₃ is D or E; α₅ is S or G; and α₆ is S, L, or T. In some embodiments,as is S, G, or R. Sequencing of individual clones isolated from theoutput of the antibody selection process revealed that R occurred atposition as at nearly the same frequency as G.

In some aspects, if α₂ is A, then α₃ is not D; α₅ is not S; or α₆ is notS; or combinations thereof.

In some aspects, α₂ is V or S; α₃ is E; as is G; or α₆ is L or T; orcombinations thereof.

In some aspects, α₂ is not A; α₃ is not D; α₅ is not S; or α₆ is not S;or combinations thereof.

In some embodiments, the antibody comprises a CDR-H3 sequence defined bythe consensus sequence ß₁-G-Y-ß₄-ß₅-Y-ß₇-β₈-F-ß₁₀-ß₁₁, where ß₁ is notpresent or Q; ß₄ is G or D; ß₅ is N or V; ß₇ is L or S; ß₈ is Y or W;ß₁₀ is D or A; and ß₁₁ is V or Y.

2.8.2. Chothia CDR-H1 Consensus Sequences

In some embodiments, the antibody comprises a Chothia CDR-H1 sequencedefined by the consensus sequence G-ε₂-ε₃-ε₄-ε₅-ε₆-ε₇, where ε₂ is Y orF; ε₃ is T, R or I; ε₄ is F or L; ε₅ is S, E, T, P, or R; ε₆ is T, S, H,Q, R, or W; and ε₇ is F, Y, or Q.

In some aspects, if ε₅ is T, then ε₆ is not S.

In some aspects, ε₅ is S, E, P, or R; or ε₆ is T, H, Q, R, or W; orcombinations thereof.

In some aspects, ε₂ is not Y; ε₃ is not T or R; ε₅ is not T; ε₆ is notS; or ε₇ is not Y; or combinations thereof.

2.8.3. Kabat CDR-H2 Consensus Sequences

In some embodiments, the antibody comprises a Kabat CDR-H2 sequencedefined by the consensus sequence W-γ₂-S-A-γ₅-N-G-N-T-γ₁₀-Y-A-Q-K-L-Q-G,where γ2 is I or V; γ5 is Y or H; and γ10 is K or N.

In some aspects, γ2 is not I; γ5 is not Y; or γ10 is not N; orcombinations thereof.

In some embodiments, the antibody comprises a Kabat CDR-H2 sequencedefined by the consensus sequenceδ₁-I-S-G-δ₅-G-δ₇-δ₈-T-Y-Y-δ₁₂-D-S-V-δ₁₆-G, where δ₁ is T or A; δ₅ is Sor G; δ₇ is S or G; δ₈ is S, D or N; δ₁₂ is A, P or S; and δ₁₆ is K orQ.

In some aspects, if δ₁ is A, then δ₅ is not S; δ₇ is not G; δ₈ is not S;δ₁₂ is not A; or δ₁₆ is not K; or combinations thereof.

In some aspects, δ₁ is T; δ₅ is G; δ₇ is S; δ₈ is D or N; or δ₁₂ is P orS; or combinations thereof.

In some aspects, δ₁ is not A; δ₅ is not S; δ₇ is not G; δ₈ is not S; δ₁₂is not A; or δ₁₆ is not K; or combinations thereof.

2.8.4. Kabat CDR-H1 Consensus Sequences

In some embodiments, the antibody comprises a Kabat CDR-H1 sequencedefined by the consensus sequence Θ₁-Θ₂-G-Θ₄-S, where Θ₁ is T, R, W, Q,H, or S; Θ₂ is Y, F, or Q; and Θ₄ is M or I.

In some aspects, Θ₁ is T, R, W, Q, or H; Θ₂ is F or Q; or Θ₄ is M; orcombinations thereof.

In some aspects, Θ₁ is not S; Θ₂ is not Y; or Θ₄ is not I; orcombinations thereof.

2.8.5. CDR-L3 Consensus Sequences

In some embodiments, the antibody comprises a CDR-L3 sequence defined bythe consensus sequence Q-Q-π₃-π₄-π₅-π₆-P-π₈-T, where π₃ is N, S, or W;π₄ is Y, K, or I; π₅ is N, E, or S; π₆ is S, V, D, or T; and π₈ is Y orW.

In some aspects, if π₄ is Y, then π₃ is not S; π₅ is not S; π₆ is not T;or π₈ is not W; or combinations thereof.

In some aspects, π₃ is N or W; π₄ is K or I; π₅ is N or E; π₆ is S, V,or D; or π₈ is Y; or combinations thereof.

In some aspects, π₃ is not S; π₄ is not Y; π₅ is not S; π₆ is not T; orπ₈ is not W; or combinations thereof.

2.8.6. CDR-L1 Consensus Sequences

In some embodiments, the antibody comprises a CDR-L1 sequence defined bythe consensus sequence S-G-D-A-L-μ6-μ7-Q-Y-μ10-Y, where μ6 is P, T, orS; μ7 is M, T, E, or K; and μ₁₀ is G or A.

In some aspects, if μ₆ is P, then μ₇ is not K, μ₁₀ is not A, orcombinations thereof.

In some aspects, μ₆ is T or S; μ₇ is M, T, or E; or μ₁₀ is G; orcombinations thereof.

In some aspects, μ₆ is not P; μ₇ is not K; or μ₁₀ is not A; orcombinations thereof.

In some embodiments, the antibody comprises a CDR-L1 sequence defined bythe consensus sequence R-A-S-E-Σ₅-V-D-Σ₈-Σ₉-G-Σ₁₁-S-F-M-Σ₁₅, where Σ₅ isS or N; Σ₈ is N or D; Σ₉ is S or Y; Σ₁₁ is I or V; and Σ₁₅ is S or N.

3. Germline

In some embodiments, the antibody that specifically binds PD-1 is anantibody comprising a variable region that is encoded by a particulargermline gene, or a variant thereof. The illustrative antibodiesprovided herein comprise variable regions that are encoded by the heavychain variable region germline genes VH1-18, VH3-21, VH3-7, and VH3-15,or variants thereof; and the light chain variable region germline genesVλ3-25, Vκ1-9, Vκ3-11, Vκ3-20, and Vκ4-1, or variants thereof. One ofskill in the art would recognize that the CDR sequences provided hereinmay also be useful when combined with variable regions encoded by othervariable region germline genes, or variants thereof. In particular, theCDR sequences provided herein may be useful when combined with variableregions encoded by variable region germline genes, or variants thereof,that are structurally similar to the variable region germline genesrecited above. For example, in some embodiments, a CDR-H sequenceprovided herein may be combined with a variable region encoded by avariable region germline gene selected from the VH1 or VH3 family, or avariant thereof. In some embodiments, a CDR-L sequence provided hereinmay be combined with a variable region encoded by a variable regiongermline gene selected from the Vλ3, Vκ1, Vκ3, and Vκ4 families, or avariant thereof.

4. Affinity

In some embodiments, the affinity of the antibody for PD-1, as indicatedby K_(D), is less than about 10⁻⁵ M, less than about 10⁻⁶ M, less thanabout 10⁻⁷ M, less than about 10⁻⁸ M, less than about 10⁻⁹ M, less thanabout 10⁻¹⁰ M, less than about 10⁻¹¹ M, or less than about 10⁻¹² M. Insome embodiments, the affinity of the antibody is between about 10⁻⁷ Mand 10⁻¹¹ M. In some embodiments, the affinity of the antibody isbetween about 10⁻⁷ M and 10⁻¹⁰ M. In some embodiments, the affinity ofthe antibody is between about 10⁻⁷ M and 10⁻⁹ M. In some embodiments,the affinity of the antibody is between about 10⁻⁷ M and 10⁻⁸ M. In someembodiments, the affinity of the antibody is between about 10⁻⁸ M and10⁻¹¹ M. In some embodiments, the affinity of the antibody is betweenabout 10⁻⁸ M and 10⁻¹⁰ M. In some embodiments, the affinity of theantibody is between about 10⁻⁹ M and 10⁻¹¹ M. In some embodiments, theaffinity of the antibody is between about 10⁻¹⁰ M and 10⁻¹¹M.

In some embodiments, the affinity of the antibody for human PD-1 isbetween about 3.85×10⁻⁸ M and 2.52×10⁻¹⁰ M. In some embodiment, theaffinity of the antibody for human PD-1 is about 2.55×10⁻⁸ M, about1.52×10⁻⁸ M, about 9.52×10⁻⁹ M, about 1.09×10⁻⁸ M, about 4.50×10⁻⁹ M,about 1.90×10⁻⁹ M, about 4.76×10⁻⁹ M, about 4.5×10⁻⁹ M, about 1.04×10⁻⁸M, about 9.90×10⁻⁹ M, about 9.13×10⁻¹⁰ M, about 2.52×10⁻¹⁰ M, about2.58×10⁻⁹ M, about 3.85×10⁻⁸ M, about 3.66×10⁻⁹ M, about 3.15×10⁻⁹ M,about 5.14×10⁻⁹ M, about 2.47×10⁻⁹M, about 2.79×10⁻⁹M, about 1.20×10⁻⁹M,or about 1.28×10⁻⁸ M

In some embodiments, the affinity of the antibody for human PD-1expressed on the surface of a cell is between about 3.2 and about 0.2nM. In some embodiment, the affinity of the antibody for human PD-1expressed on the surface of a cell is about 0.2 nM, about 0.4 nM, about0.9 nM, about 1 nM, about 0.3 nM, about 0.7 nM, about 0.2 nM, about 0.8nM, about 3.2 nM, about 2.9 nM, about 1.39 nM, or about 1.34 nM.

In some embodiments, the affinity of the antibody for murine PD-1 isbetween about 6.09×10⁻⁸ M and 9.08×10⁻⁹ M. In some embodiment, theaffinity of the antibody for murine PD-1 is about 6.09×10⁻⁸ M, about6.22×10⁻⁸ M, or about 9.08×10⁻⁹ M.

In some embodiments, the affinity of the antibody for cynomolgus PD-1 isbetween about 2.43×10⁻⁸ M and 1.95×10⁻¹⁰ M. In some embodiment, theaffinity of the antibody for cynomolgus PD-1 is about 2.43×10⁻⁸ M, about1.55×10⁻⁸ M, about 2.22×10⁻⁸ M, about 2.56×10⁻⁹M, about 2.54×10⁻⁹M,about 5.61×10⁻¹⁰ M, or about 1.95×10⁻¹⁰ M

In some embodiments the antibody has a k_(a) of at least about 10⁴M⁻¹×sec⁻¹. In some embodiments the antibody has a k_(a) of at leastabout 10⁵ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_(a) of atleast about 10⁶ M⁻¹×sec⁻¹. In some embodiments the antibody has a k_(a)of between about 10⁴ M⁻¹×sec⁻¹ and about 10⁵ M⁻¹×sec⁻¹. In someembodiments the antibody has a k_(a) of between about 10⁵ M⁻¹×sec⁻¹ andabout 10⁶ M⁻¹×sec⁻¹.

In some embodiments the antibody has a k_(a) when associating with humanPD-1 of between about 4.74×10⁴ M⁻¹×sec⁻¹ and about 1.23×10⁶ M⁻¹×sec⁻¹.In some embodiments the antibody has a k_(a) when associating with humanPD-1 of about 4.88×10⁵ M⁻¹×sec⁻¹, about 1.23×10⁶ M⁻¹×sec⁻¹, about7.37×10⁵ M⁻¹×sec⁻¹, about 6.87×10⁵ M⁻¹×sec⁻¹, about 5.63×10⁵ M⁻¹×sec⁻¹,about 5.16×10⁵ M⁻¹×sec⁻¹, about 2.48×10⁵ M⁻¹×sec⁻¹, about 7.98×10⁵M⁻¹×sec⁻¹, about 1.82×10⁵ M⁻¹×sec⁻¹, about 4.74×10⁴ M⁻¹×sec⁻¹, about1.85×10⁵ M⁻¹×sec⁻¹, about 2.00×10⁵ M⁻¹×sec⁻¹, about 8.12×10⁴ M⁻¹×sec⁻¹,about 1.21×10⁶ M⁻¹×sec⁻¹, about 1.16×10⁶ M⁻¹×sec⁻¹, about 5.13×10⁵M⁻¹×sec⁻¹, or about 1.86×10⁵ M⁻¹×sec⁻¹.

In some embodiments the antibody has a k_(d) of about 10⁻⁵ sec⁻¹ orless. In some embodiments the antibody has a k_(d) of about 10⁴ sec⁻¹ orless. In some embodiments the antibody has a k_(d) of about 10⁻³ sec⁻¹or less. In some embodiments the antibody has a k_(d) of between about10⁻² sec⁻¹ and about 10⁻⁵ sec⁻¹. In some embodiments the antibody has ak_(d) of between about 10⁻² sec⁻¹ and about 10⁻⁴ sec⁻¹. In someembodiments the antibody has a k_(d) of between about 10⁻³ sec⁻¹ andabout 10⁻⁵ sec⁻¹.

In some embodiments the antibody has a k_(d) when dissociating fromhuman PD-1 of between about 1.87×10⁻² sec⁻¹ and about 4.17×10⁻⁴ sec⁻¹.In some embodiments the antibody has a k_(d) when dissociating fromhuman PD-1 of about 1.24×10⁻² sec⁻¹, about 1.87×10⁻² sec⁻¹, about7.01×10⁻³ sec⁻¹, about 7.74×10⁻³ sec⁻¹, about 2.54×10⁻³ sec⁻¹, about9.80×10⁻⁴ sec⁻¹, about 1.18×10⁻³ sec⁻¹, about 3.59×10⁻³ sec⁻¹, about4.68×10⁻⁴ sec⁻¹, about 1.82×10⁻³ sec⁻¹, about 6.79×10⁻⁴ sec⁻¹, about6.28×10⁻⁴ sec⁻¹, about 4.17×10⁻⁴ sec⁻¹, about 2.99×10⁻³ sec⁻¹, about3.24×10⁻³ sec⁻¹, about 6.17×10⁻⁴ sec⁻¹, or about 2.39×10⁻³ sec⁻¹.

In some aspects, the K_(D), k_(a), and k_(d) are determined at 25° C. Insome embodiments, the K_(D), k_(a), and k_(d) are determined by surfaceplasmon resonance. In some embodiments, the K_(D), k_(a), and k_(d) aredetermined according to the methods described in Examples 4 and 6.

5. Inhibition of PD-L1 and PD-L2 Binding

In some embodiments, the antibody inhibits binding of one or more ofPD-L1 and PD-L2 to PD-1.

In some embodiments, the antibody inhibits binding of PD-L1 to PD-1 withan IC50 of about 1 to about 7 nM. In some aspects, the antibody inhibitsbinding of PD-L1 to PD-1 with an IC50 of about 1.99, about 2.53, about5.86, or about 5.96 nM.

In some embodiments, the antibody inhibits binding of PD-L2 to PD-1 withan IC₅₀ of about 0.01 to about 1 nM. In some aspects, the antibodyinhibits binding of PD-L2 to PD-1 with an IC₅₀ of about 0.01, about0.18, about 0.56, or about 0.58 nM.

In some aspects, the antibody inhibits binding of PD-L1 to PD-1 with anIC₅₀ of about 5.96 nM, and inhibits binding of PD-L2 to PD-1 with anIC₅₀ of about 0.56 nM. In some aspects, the antibody inhibits binding ofPD-L1 to PD-1 with an IC₅₀ of about 5.86 nM, and inhibits binding ofPD-L2 to PD-1 with an IC₅₀ of about 0.58 nM. In some aspects, theantibody inhibits binding of PD-L1 to PD-1 with an IC₅₀ of about 1.99nM, and inhibits binding of PD-L2 to PD-1 with an IC₅₀ of about 0.01 nM.In some aspects, the antibody inhibits binding of PD-L1 to PD-1 with anIC₅₀ of about 2.53 nM, and inhibits binding of PD-L2 to PD-1 with anIC₅₀ of about 0.18 nM.

6. PD-1 Assays

In some embodiments, the anti-PD-1 antibodies induce the secretion ofinterferon gamma when added to a peripheral blood mononuclear cell(PBMC) two-way mixed lymphocyte reaction (MLR) assay, as described inExamples 8 and 16.

In some embodiments, the anti-PD-1 antibodies induce the secretion ofinterferon gamma when added to a PBMC cytomegalovirus recall assay, asdescribed in Example 16.

In some embodiments, the anti-PD-1 antibodies accelerate the onset ofgraft versus host disease, as shown in Example 18.

7. Glycosylation Variants

In certain embodiments, an antibody may be altered to increase, decreaseor eliminate the extent to which it is glycosylated. Glycosylation ofpolypeptides is typically either “N-linked” or “O-linked.”

“N-linked” glycosylation refers to the attachment of a carbohydratemoiety to the side chain of an asparagine residue. The tripeptidesequences asparagine-X-serine and asparagine-X-threonine, where X is anyamino acid except proline, are the recognition sequences for enzymaticattachment of the carbohydrate moiety to the asparagine side chain.Thus, the presence of either of these tripeptide sequences in apolypeptide creates a potential glycosylation site.

“O-linked” glycosylation refers to the attachment of one of the sugarsN-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, mostcommonly serine or threonine, although 5-hydroxyproline or5-hydroxylysine may also be used.

Addition or deletion of N-linked glycosylation sites to the antibody maybe accomplished by altering the amino acid sequence such that one ormore of the above-described tripeptide sequences is created or removed.Addition or deletion of O-linked glycosylation sites may be accomplishedby addition, deletion, or substitution of one or more serine orthreonine residues in or to (as the case may be) the sequence of anantibody.

8. Fc Variants

In certain embodiments, amino acid modifications may be introduced intothe Fc region of an antibody provided herein to generate an Fc regionvariant. In certain embodiments, the Fc region variant possesses some,but not all, effector functions. Such antibodies may be useful, forexample, in applications in which the half-life of the antibody in vivois important, yet certain effector functions are unnecessary ordeleterious. Examples of effector functions include complement-dependentcytotoxicity (CDC) and antibody-directed complement-mediatedcytotoxicity (ADCC). Numerous substitutions or substitutions ordeletions with altered effector function are known in the art.

An alteration in in CDC and/or ADCC activity can be confirmed using invitro and/or in vivo assays. For example, Fc receptor (FcR) bindingassays can be conducted to measure FcγR binding. The primary cells formediating ADCC, NK cells, express FcγRIII only, whereas monocytesexpress FcγRI, FcγRII and FcγRIII. FcR expression on hematopoietic cellsis summarized in Ravetch and Kinet, Ann. Rev. Immunol., 1991, 9:457-492.

Non-limiting examples of in vitro assays to assess ADCC activity of amolecule of interest are provided in U.S. Pat. Nos. 5,500,362 and5,821,337; Hellstrom et al., Proc. Natl. Acad. Sci. USA., 1986,83:7059-7063; Hellstrom et al., Proc. Natl. Acad. Sci. USA., 1985,82:1499-1502; and Bruggemann et al., J. Exp. Med., 1987, 166:1351-1361.Useful effector cells for such assays include peripheral bloodmononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively,or additionally, ADCC activity of the molecule of interest may beassessed in vivo, using an animal model such as that disclosed in Clyneset al. Proc. Natl. Acad. Sci. USA., 1998, 95:652-656.

C1q binding assays may also be carried out to confirm that the antibodyis unable to bind C1q and hence lacks CDC activity. Examples of C1qbinding assays include those described in WO 2006/029879 and WO2005/100402.

Complement activation assays include those described, for example, inGazzano-Santoro et al., J. Immunol. Methods, 1996, 202:163-171; Cragg etal., Blood, 2003, 101:1045-1052; and Cragg and Glennie, Blood, 2004,103:2738-2743.

FcRn binding and in vivo clearance (half-life determination) can also bemeasured, for example, using the methods described in Petkova et al.,Intl. Immunol., 2006, 18:1759-1769.

9. Preparation of Antibodies

9.1. Antigen Preparation

The PD-1 antigen to be used for production of antibodies may be intactPD-1 or a fragment of PD-1. The intact PD-1, or fragment of PD-1, may bein the form of an isolated protein or expressed by a cell. Other formsof PD-1 useful for generating antibodies will be apparent to thoseskilled in the art.

9.2. Monoclonal Antibodies

Monoclonal antibodies may be obtained, for example, using the hybridomamethod first described by Kohler et al., Nature, 1975, 256:495-497,and/or by recombinant DNA methods (see e.g., U.S. Pat. No. 4,816,567).Monoclonal antibodies may also be obtained, for example, using phage oryeast-based libraries. See e.g., U.S. Pat. Nos. 8,258,082 and 8,691,730.

In the hybridoma method, a mouse or other appropriate host animal isimmunized to elicit lymphocytes that produce or are capable of producingantibodies that will specifically bind to the protein used forimmunization. Alternatively, lymphocytes may be immunized in vitro.Lymphocytes are then fused with myeloma cells using a suitable fusingagent, such as polyethylene glycol, to form a hybridoma cell. See GodingJ. W., Monoclonal Antibodies: Principles and Practice 3^(rd) ed. (1986)Academic Press, San Diego, Calif.

The hybridoma cells are seeded and grown in a suitable culture mediumthat contains one or more substances that inhibit the growth or survivalof the unfused, parental myeloma cells. For example, if the parentalmyeloma cells lack the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT), the culture medium for the hybridomastypically will include hypoxanthine, aminopterin, and thymidine (HATmedium), which substances prevent the growth of HGPRT-deficient cells.

Useful myeloma cells are those that fuse efficiently, support stablehigh-level production of antibody by the selected antibody-producingcells, and are sensitive media conditions, such as the presence orabsence of HAT medium. Among these, preferred myeloma cell lines aremurine myeloma lines, such as those derived from MOP-21 and MC-11 mousetumors (available from the Salk Institute Cell Distribution Center, SanDiego, Calif.), and SP-2 or X63-Ag8-653 cells (available from theAmerican Type Culture Collection, Rockville, Md.). Human myeloma andmouse-human heteromyeloma cell lines also have been described for theproduction of human monoclonal antibodies. See e.g., Kozbor, J.Immunol., 1984, 133:3001.

After the identification of hybridoma cells that produce antibodies ofthe desired specificity, affinity, and/or biological activity, selectedclones may be subcloned by limiting dilution procedures and grown bystandard methods. See Goding, supra. Suitable culture media for thispurpose include, for example, D-MEM or RPMI-1640 medium. In addition,the hybridoma cells may be grown in vivo as ascites tumors in an animal.

DNA encoding the monoclonal antibodies may be readily isolated andsequenced using conventional procedures (e.g., by using oligonucleotideprobes that are capable of binding specifically to genes encoding theheavy and light chains of the monoclonal antibodies). Thus, thehybridoma cells can serve as a useful source of DNA encoding antibodieswith the desired properties. Once isolated, the DNA may be placed intoexpression vectors, which are then transfected into host cells such asbacteria (e.g., E. coli), yeast (e.g., Saccharomyces or Pichia sp.), COScells, Chinese hamster ovary (CHO) cells, or myeloma cells that do nototherwise produce antibody, to produce the monoclonal antibodies.

9.3. Humanized Antibodies

Humanized antibodies may be generated by replacing most, or all, of thestructural portions of a monoclonal antibody with corresponding humanantibody sequences. Consequently, a hybrid molecule is generated inwhich only the antigen-specific variable, or CDR, is composed ofnon-human sequence. Methods to obtain humanized antibodies include thosedescribed in, for example, Winter and Milstein, Nature, 1991,349:293-299; Rader et al., Proc. Nat. Acad. Sci. USA., 1998,95:8910-8915; Steinberger et al., J. Biol. Chem., 2000, 275:36073-36078;Queen et al., Proc. Natl. Acad. Sci. USA., 1989, 86:10029-10033; andU.S. Pat. Nos. 5,585,089, 5,693,761, 5,693,762, and 6,180,370.

9.4. Human Antibodies

Human antibodies can be generated by a variety of techniques known inthe art, for example by using transgenic animals (e.g., humanized mice).See, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA., 1993,90:2551; Jakobovits et al., Nature, 1993, 362:255-258; Bruggermann etal., Year in Immuno., 1993, 7:33; and U.S. Pat. Nos. 5,591,669,5,589,369 and 5,545,807. Human antibodies can also be derived fromphage-display libraries (see e.g., Hoogenboom et al., J. Mol. Biol.,1991, 227:381-388; Marks et al., J. Mol. Biol., 1991, 222:581-597; andU.S. Pat. Nos. 5,565,332 and 5,573,905). Human antibodies may also begenerated by in vitro activated B cells (see e.g., U.S. Pat. Nos.5,567,610 and 5,229,275). Human antibodies may also be derived fromyeast-based libraries (see e.g., U.S. Pat. No. 8,691,730).

10. Vectors, Host Cells, and Recombinant Methods

The invention also provides isolated nucleic acids encoding anti-PD-1antibodies, vectors and host cells comprising the nucleic acids, andrecombinant techniques for the production of the antibodies.

For recombinant production of the antibody, the nucleic acid encoding itmay be isolated and inserted into a replicable vector for furthercloning (i.e., amplification of the DNA) or expression. In some aspects,the nucleic acid may be produced by homologous recombination, forexample as described in U.S. Pat. No. 5,204,244.

Many different vectors are known in the art. The vector componentsgenerally include, but are not limited to, one or more of the following:a signal sequence, an origin of replication, one or more marker genes,an enhancer element, a promoter, and a transcription terminationsequence, for example as described in U.S. Pat. No. 5,534,615.

Illustrative examples of suitable host cells are provided below, thesehost cells are not meant to be limiting.

Suitable host cells include any prokaryotic (e.g., bacterial), lowereukaryotic (e.g., yeast), or higher eukaryotic (e.g., mammalian) cells.Suitable prokaryotes include eubacteria, such as Gram-negative orGram-positive organisms, for example, Enterobacteriaceae such asEscherichia (E. coli), Enterobacter, Erwinia, Klebsiella, Proteus,Salmonella (S. typhimurium), Serratia (S. marcescans), Shigella, Bacilli(B. subtilis and B. licheniformis), Pseudomonas (P. aeruginosa), andStreptomyces. One useful E. coli cloning host is E. coli 294, althoughother strains such as E. coli B, E. coli X1776, and E. coli W3110 aresuitable.

In addition to prokaryotes, eukaryotic microbes such as filamentousfungi or yeast are also suitable cloning or expression hosts foranti-PD-1 antibody-encoding vectors. Saccharomyces cerevisiae, or commonbaker's yeast, is a commonly used lower eukaryotic host microorganism.However, a number of other genera, species, and strains are availableand useful, such as Schizosaccharomyces pombe, Kluyveromyces (K. lactis,K. fragilis, K. bulgaricus K. wickeramii, K. waltii, K. drosophilarum,K. thermotolerans, and K. marxianus), Yarrowia, Pichia pastoris, Candida(C. albicans), Trichoderma reesia, Neurospora crassa, Schwanniomyces (S.occidentalis), and filamentous fungi such as, for example Penicillium,Tolypocladium, and Aspergillus (A. nidulans and A. niger).

Useful mammalian host cells include COS-7 cells, HEK293 cells; babyhamster kidney (BHK) cells; Chinese hamster ovary (CHO); mouse sertolicells; African green monkey kidney cells (VERO-76), and the like.

The host cells used to produce the anti-PD-1 antibody of this inventionmay be cultured in a variety of media. Commercially available media suchas, for example, Ham's F10, Minimal Essential Medium (MEM), RPMI-1640,and Dulbecco's Modified Eagle's Medium (DMEM) are suitable for culturingthe host cells. In addition, any of the media described in Ham et al.,Meth. Enz., 1979, 58:44; Barnes et al., Anal. Biochem., 1980, 102:255;and U.S. Pat. Nos. 4,767,704, 4,657,866, 4,927,762, 4,560,655, and5,122,469, or WO 90/03430 and WO 87/00195 may be used.

Any of these media may be supplemented as necessary with hormones and/orother growth factors (such as insulin, transferrin, or epidermal growthfactor), salts (such as sodium chloride, calcium, magnesium, andphosphate), buffers (such as HEPES), nucleotides (such as adenosine andthymidine), antibiotics, trace elements (defined as inorganic compoundsusually present at final concentrations in the micromolar range), andglucose or an equivalent energy source. Any other necessary supplementsmay also be included at appropriate concentrations that would be knownto those skilled in the art.

The culture conditions, such as temperature, pH, and the like, are thosepreviously used with the host cell selected for expression, and will beapparent to the ordinarily skilled artisan.

When using recombinant techniques, the antibody can be producedintracellularly, in the periplasmic space, or directly secreted into themedium. If the antibody is produced intracellularly, as a first step,the particulate debris, either host cells or lysed fragments, isremoved, for example, by centrifugation or ultrafiltration. For example,Carter et al. (Bio/Technology, 1992, 10:163-167) describes a procedurefor isolating antibodies which are secreted to the periplasmic space ofE. coli. Briefly, cell paste is thawed in the presence of sodium acetate(pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30min. Cell debris can be removed by centrifugation.

In some embodiments, the antibody is produced in a cell-free system. Insome aspects, the cell-free system is an in vitro transcription andtranslation system as described in Yin et al., mAbs, 2012, 4:217-225,incorporated by reference in its entirety. In some aspects, thecell-free system utilizes a cell-free extract from a eukaryotic cell orfrom a prokaryotic cell. In some aspects, the prokaryotic cell is E.coli. Cell-free expression of the antibody may be useful, for example,where the antibody accumulates in a cell as an insoluble aggregate, orwhere yields from periplasmic expression are low.

Where the antibody is secreted into the medium, supernatants from suchexpression systems are generally first concentrated using a commerciallyavailable protein concentration filter, for example, an Amicon® orMillipore® Pellcon® ultrafiltration unit. A protease inhibitor such asPMSF may be included in any of the foregoing steps to inhibitproteolysis and antibiotics may be included to prevent the growth ofadventitious contaminants.

The antibody composition prepared from the cells can be purified using,for example, hydroxylapatite chromatography, gel electrophoresis,dialysis, and affinity chromatography, with affinity chromatographybeing a particularly useful purification technique. The suitability ofprotein A as an affinity ligand depends on the species and isotype ofany immunoglobulin Fc domain that is present in the antibody. Protein Acan be used to purify antibodies that are based on human γ₁, γ₂, or γ₄heavy chains (Lindmark et al., J. Immunol. Meth., 1983, 62:1-13).Protein G is useful for all mouse isotypes and for human γ3 (Guss etal., EMBO J., 1986, 5:1567-1575).

The matrix to which the affinity ligand is attached is most oftenagarose, but other matrices are available. Mechanically stable matricessuch as controlled pore glass or poly(styrenedivinyl)benzene allow forfaster flow rates and shorter processing times than can be achieved withagarose. Where the antibody comprises a C_(H3) domain, the BakerBondABX® resin is useful for purification.

Other techniques for protein purification, such as fractionation on anion-exchange column, ethanol precipitation, Reverse Phase HPLC,chromatography on silica, chromatography on heparin Sepharose®,chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are alsoavailable, and can be applied by one of skill in the art.

Following any preliminary purification step(s), the mixture comprisingthe antibody of interest and contaminants may be subjected to low pHhydrophobic interaction chromatography using an elution buffer at a pHbetween about 2.5 to about 4.5, generally performed at low saltconcentrations (e.g., from about 0 to about 0.25 M salt).

11. Pharmaceutical Compositions and Methods of Administration

Any of the antibodies provided herein can be provided in any appropriatepharmaceutical composition and be administered by any suitable route ofadministration. Suitable routes of administration include, but are notlimited to, the inhalation, intraarterial, intradermal, intramuscular,intraperitoneal, intravenous, nasal, parenteral, pulmonary, andsubcutaneous routes.

The pharmaceutical composition may comprise one or more pharmaceuticalexcipients. Any suitable pharmaceutical excipient may be used, and oneof ordinary skill in the art is capable of selecting suitablepharmaceutical excipients. Accordingly, the pharmaceutical excipientsprovided below are intended to be illustrative, and not limiting.Additional pharmaceutical excipients include, for example, thosedescribed in the Handbook of Pharmaceutical Excipients, Rowe et al.(Eds.) 6th Ed. (2009), incorporated by reference in its entirety.

In some embodiments, the pharmaceutical composition comprises ananti-foaming agent. Any suitable anti-foaming agent may be used. In someaspects, the anti-foaming agent is selected from an alcohol, an ether,an oil, a wax, a silicone, a surfactant, and combinations thereof. Insome aspects, the anti-foaming agent is selected from a mineral oil, avegetable oil, ethylene bis stearamide, a paraffin wax, an ester wax, afatty alcohol wax, a long chain fatty alcohol, a fatty acid soap, afatty acid ester, a silicon glycol, a fluorosilicone, a polyethyleneglycol-polypropylene glycol copolymer, polydimethylsiloxane-silicondioxide, ether, octyl alcohol, capryl alcohol, sorbitan trioleate, ethylalcohol, 2-ethyl-hexanol, dimethicone, oleyl alcohol, simethicone, andcombinations thereof.

In some embodiments, the pharmaceutical composition comprises acosolvent. Illustrative examples of cosolvents include ethanol,poly(ethylene) glycol, butylene glycol, dimethylacetamide, glycerin, andpropylene glycol.

In some embodiments, the pharmaceutical composition comprises a buffer.Illustrative examples of buffers include acetate, borate, carbonate,lactate, malate, phosphate, citrate, hydroxide, diethanolamine,monoethanolamine, glycine, methionine, guar gum, and monosodiumglutamate.

In some embodiments, the pharmaceutical composition comprises a carrieror filler. Illustrative examples of carriers or fillers include lactose,maltodextrin, mannitol, sorbitol, chitosan, stearic acid, xanthan gum,and guar gum.

In some embodiments, the pharmaceutical composition comprises asurfactant. Illustrative examples of surfactants include d-alphatocopherol, benzalkonium chloride, benzethonium chloride, cetrimide,cetylpyridinium chloride, docusate sodium, glyceryl behenate, glycerylmonooleate, lauric acid, macrogol 15 hydroxystearate, myristyl alcohol,phospholipids, polyoxyethylene alkyl ethers, polyoxyethylene sorbitanfatty acid esters, polyoxyethylene stearates, polyoxylglycerides, sodiumlauryl sulfate, sorbitan esters, and vitamin E polyethylene(glycol)succinate.

In some embodiments, the pharmaceutical composition comprises ananti-caking agent. Illustrative examples of anti-caking agents includecalcium phosphate (tribasic), hydroxymethyl cellulose, hydroxypropylcellulose, and magnesium oxide.

Other excipients that may be used with the pharmaceutical compositionsinclude, for example, albumin, antioxidants, antibacterial agents,antifungal agents, bioabsorbable polymers, chelating agents, controlledrelease agents, diluents, dispersing agents, dissolution enhancers,emulsifying agents, gelling agents, ointment bases, penetrationenhancers, preservatives, solubilizing agents, solvents, stabilizingagents, and sugars. Specific examples of each of these agents aredescribed, for example, in the Handbook of Pharmaceutical Excipients,Rowe et al. (Eds.) 6th Ed. (2009), The Pharmaceutical Press,incorporated by reference in its entirety.

In some embodiments, the pharmaceutical composition comprises a solvent.In some aspects, the solvent is saline solution, such as a sterileisotonic saline solution or dextrose solution. In some aspects, thesolvent is water for injection.

In some embodiments, the pharmaceutical compositions are in aparticulate form, such as a microparticle or a nanoparticle.Microparticles and nanoparticles may be formed from any suitablematerial, such as a polymer or a lipid. In some aspects, themicroparticles or nanoparticles are micelles, liposomes, orpolymersomes. In certain embodiments, a composition provided herein is apharmaceutical composition or a single unit dosage form. Pharmaceuticalcompositions and single unit dosage forms provided herein comprise aprophylactically or therapeutically effective amount of one or moreprophylactic or therapeutic antibodies.

Further encompassed herein are anhydrous pharmaceutical compositions anddosage forms comprising an antibody, since water can facilitate thedegradation of some antibodies.

Anhydrous pharmaceutical compositions and dosage forms provided hereincan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine can be anhydrousif substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions can be packaged using materials known to prevent exposureto water such that they can be included in suitable formulary kits.Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

11.1. Parenteral Dosage Forms

In certain embodiments, provided are parenteral dosage forms. Parenteraldosage forms can be administered to subjects by various routesincluding, but not limited to, subcutaneous, intravenous (includingbolus injection), intramuscular, and intraarterial. Because theiradministration typically bypasses subjects' natural defenses againstcontaminants, parenteral dosage forms are typically, sterile or capableof being sterilized prior to administration to a subject. Examples ofparenteral dosage forms include, but are not limited to, solutions readyfor injection, dry products ready to be dissolved or suspended in apharmaceutically acceptable vehicle for injection, suspensions ready forinjection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage formsare well known to those skilled in the art. Examples include, but arenot limited to: Water for Injection USP; aqueous vehicles such as, butnot limited to, Sodium Chloride Injection, Ringer's Injection, DextroseInjection, Dextrose and Sodium Chloride Injection, and Lactated Ringer'sInjection; water miscible vehicles such as, but not limited to, ethylalcohol, polyethylene glycol, and polypropylene glycol; and non-aqueousvehicles such as, but not limited to, corn oil, cottonseed oil, peanutoil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

Excipients that increase the solubility of one or more of the antibodiesdisclosed herein can also be incorporated into the parenteral dosageforms.

11.2. Dosage and Unit Dosage Forms

In human therapeutics, the doctor will determine the posology which heconsiders most appropriate according to a preventive or curativetreatment and according to the age, weight, condition and other factorsspecific to the subject to be treated.

The amount of the antibody or composition which will be effective in theprevention or treatment of a disorder or one or more symptoms thereofwill vary with the nature and severity of the disease or condition, andthe route by which the antibody is administered. The frequency anddosage will also vary according to factors specific for each subjectdepending on the specific therapy (e.g., therapeutic or prophylacticagents) administered, the severity of the disorder, disease, orcondition, the route of administration, as well as age, body, weight,response, and the past medical history of the subject. Effective dosesmay be extrapolated from dose-response curves derived from in vitro oranimal model test systems.

In certain embodiments, exemplary doses of a composition includemilligram or microgram amounts of the antibody per kilogram of subjector sample weight (e.g., about 10 micrograms per kilogram to about 50milligrams per kilogram, about 100 micrograms per kilogram to about 25milligrams per kilogram, or about 100 microgram per kilogram to about 10milligrams per kilogram). In certain embodiment, the dosage of theantibody provided herein, based on weight of the antibody, administeredto prevent, treat, manage, or ameliorate a disorder, or one or moresymptoms thereof in a subject is 0.1 mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4mg/kg, 5 mg/kg, 6 mg/kg, 10 mg/kg, or 15 mg/kg or more of a subject'sbody weight. In another embodiment, the dosage of the composition or acomposition provided herein administered to prevent, treat, manage, orameliorate a disorder, or one or more symptoms thereof in a subject is0.1 mg to 200 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg,0.1 mg to 20 mg, 0.1 mg to 15 mg, 0.1 mg to 10 mg, 0.1 mg to 7.5 mg, 0.1mg to 5 mg, 0.1 to 2.5 mg, 0.25 mg to 20 mg, 0.25 to 15 mg, 0.25 to 12mg, 0.25 to 10 mg, 0.25 mg to 7.5 mg, 0.25 mg to 5 mg, 0.25 mg to 2.5mg, 0.5 mg to 20 mg, 0.5 to 15 mg, 0.5 to 12 mg, 0.5 to 10 mg, 0.5 mg to7.5 mg, 0.5 mg to 5 mg, 0.5 mg to 2.5 mg, 1 mg to 20 mg, 1 mg to 15 mg,1 mg to 12 mg, 1 mg to 10 mg, 1 mg to 7.5 mg, 1 mg to 5 mg, or 1 mg to2.5 mg.

The dose can be administered according to a suitable schedule, forexample, once, two times, three times, or for times weekly. It may benecessary to use dosages of the antibody outside the ranges disclosedherein in some cases, as will be apparent to those of ordinary skill inthe art. Furthermore, it is noted that the clinician or treatingphysician will know how and when to interrupt, adjust, or terminatetherapy in conjunction with subject response.

Different therapeutically effective amounts may be applicable fordifferent diseases and conditions, as will be readily known by those ofordinary skill in the art. Similarly, amounts sufficient to prevent,manage, treat or ameliorate such disorders, but insufficient to cause,or sufficient to reduce, adverse effects associated with the antibodiesprovided herein are also encompassed by the herein described dosageamounts and dose frequency schedules. Further, when a subject isadministered multiple dosages of a composition provided herein, not allof the dosages need be the same. For example, the dosage administered tothe subject may be increased to improve the prophylactic or therapeuticeffect of the composition or it may be decreased to reduce one or moreside effects that a particular subject is experiencing.

In certain embodiments, treatment or prevention can be initiated withone or more loading doses of an antibody or composition provided hereinfollowed by one or more maintenance doses.

In certain embodiments, a dose of an antibody or composition providedherein can be administered to achieve a steady-state concentration ofthe antibody in blood or serum of the subject. The steady-stateconcentration can be determined by measurement according to techniquesavailable to those of skill or can be based on the physicalcharacteristics of the subject such as height, weight and age.

In certain embodiments, administration of the same composition may berepeated and the administrations may be separated by at least 1 day, 2days, 3 days, 5 days, 10 days, 15 days, 30 days, 45 days, 2 months, 75days, 3 months, or 6 months. In other embodiments, administration of thesame prophylactic or therapeutic agent may be repeated and theadministration may be separated by at least 1 day, 2 days, 3 days, 5days, 10 days, 15 days, 30 days, 45 days, 2 months, 75 days, 3 months,or 6 months.

12. Therapeutic Applications

For therapeutic applications, the antibodies of the invention areadministered to a mammal, generally a human, in a pharmaceuticallyacceptable dosage form such as those known in the art and thosediscussed above. For example, the antibodies of the invention may beadministered to a human intravenously as a bolus or by continuousinfusion over a period of time, by intramuscular, intraperitoneal,intra-cerebrospinal, subcutaneous, intra-articular, intrasynovial,intrathecal, or intratumoral routes. The antibodies also are suitablyadministered by peritumoral, intralesional, or perilesional routes, toexert local as well as systemic therapeutic effects. The intraperitonealroute may be particularly useful, for example, in the treatment ofovarian tumors.

The antibodies provided herein may be useful for the treatment of anydisease or condition involving PD-1, such as cancer, autoimmune disease,and infection.

Any suitable cancer may be treated with the antibodies provided herein.Illustrative suitable cancers include, for example, acute lymphoblasticleukemia (ALL), acute myeloid leukemia (AML), adrenocortical carcinoma,anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, braintumor, bile duct cancer, bladder cancer, bone cancer, breast cancer,bronchial tumor, Burkitt Lymphoma, carcinoma of unknown primary origin,cardiac tumor, cervical cancer, chordoma, chronic lymphocytic leukemia(CLL), chronic myelogenous leukemia (CML), chronic myeloproliferativeneoplasm, colon cancer, colorectal cancer, craniopharyngioma, cutaneousT-cell lymphoma, ductal carcinoma, embryonal tumor, endometrial cancer,ependymoma, esophageal cancer, esthesioneuroblastoma, fibroushistiocytoma, Ewing sarcoma, eye cancer, germ cell tumor, gallbladdercancer, gastric cancer, gastrointestinal carcinoid tumor,gastrointestinal stromal tumor, gestational trophoblastic disease,glioma, head and neck cancer, hairy cell leukemia, hepatocellularcancer, histiocytosis, Hodgkin lymphoma, hypopharyngeal cancer,intraocular melanoma, islet cell tumor, Kaposi sarcoma, kidney cancer,Langerhans cell histiocytosis, laryngeal cancer, leukemia, lip and oralcavity cancer, liver cancer, lobular carcinoma in situ, lung cancer,lymphoma, macroglobulinemia, malignant fibrous histiocytoma, melanoma,Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancerwith occult primary, midline tract carcinoma involving NUT gene, mouthcancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosisfungoides, myelodysplastic syndrome, myelodysplastic/myeloproliferativeneoplasm, nasal cavity and par nasal sinus cancer, nasopharyngealcancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer,oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer,papillomatosis, paraganglioma, parathyroid cancer, penile cancer,pharyngeal cancer, pheochromocytomas, pituitary tumor, pleuropulmonaryblastoma, primary central nervous system lymphoma, prostate cancer,rectal cancer, renal cell cancer, renal pelvis and ureter cancer,retinoblastoma, rhabdoid tumor, salivary gland cancer, Sezary syndrome,skin cancer, small cell lung cancer, small intestine cancer, soft tissuesarcoma, spinal cord tumor, stomach cancer, T-cell lymphoma, teratoidtumor, testicular cancer, throat cancer, thymoma and thymic carcinoma,thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, vulvarcancer, and Wilms tumor.

Any suitable autoimmune disease may be treated with the antibodiesprovided herein. Illustrative suitable autoimmune diseases, or diseaseswith an autoimmune component, include, for example, acute disseminatedencephalomyelitis (ADEM), acute necrotizing hemorrhagicleukoencephalitis, Addison's disease, agammaglobulinemia, alopeciaareata, amyloidosis, ankylosing spondylitis, anti-GBM/anti-TBMnephritis, antiphospholipid syndrome (APS), autoimmune angioedema,autoimmune aplastic anemia, autoimmune dysautonomia, autoimmunehepatitis, autoimmune hyperlipidemia, autoimmune immunodeficiency,autoimmune inner ear disease (AIED), autoimmune myocarditis, autoimmuneoophoritis, autoimmune pancreatitis, autoimmune retinopathy, autoimmunethrombocytopenic purpura (ATP), autoimmune thyroid disease, autoimmuneurticarial, axonal & neuronal neuropathies, Balo disease, Behcet'sdisease, bullous pemphigoid, cardiomyopathy, Castleman disease, Celiacdisease, Chagas disease, chronic fatigue syndrome, chronic inflammatorydemyelinating polyneuropathy (CIDP), chronic recurrent multifocalostomyelitis (CRMO), Churg-Strauss syndrome, cicatricialpemphigoid/benign mucosal pemphigoid, Crohn's disease, Cogans syndrome,cold agglutinin disease, congenital heart block, coxsackie myocarditis,CREST disease, essential mixed cryoglobulinemia, demyelinatingneuropathies, dermatitis herpetiformis, dermatomyositis, Devic's disease(neuromyelitis optica), discoid lupus, Dressler's syndrome,endometriosis, eosinophilic esophagitis, eosinophilic fasciitis,erythema nodosum, experimental allergic encephalomyelitis, Evanssyndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis(temporal arteritis), giant cell myocarditis, glomerulonephritis,Goodpasture's syndrome, granulomatosis with polyangiitis (GPA) (formerlycalled Wegener's Granulomatosis), Graves' disease, Guillain-Barresyndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, hemolyticanemia, Henoch-Schonlein purpura, herpes gestationis,hypogammaglobulinemia, idiopathic thrombocytopenic purpura (ITP), IgAnephropathy, IgG4-related sclerosing disease, immunoregulatorylipoproteins, inclusion body myositis, interstitial cystitis, juvenilearthritis, juvenile diabetes (Type 1 diabetes), juvenile myositis,Kawasaki syndrome, Lambert-Eaton syndrome, leukocytoclastic vasculitis,lichen planus, lichen sclerosus, ligneous conjunctivitis, linear IgAdisease (LAD), lupus (SLE), Lyme disease (chronic), Meniere's disease,microscopic polyangiitis, mixed connective tissue disease (MCTD),Mooren's ulcer, Mucha-Habermann disease, multiple sclerosis, myastheniagravis, myositis, narcolepsy, neuromyelitis optica (Devic's),neutropenia, ocular cicatricial pemphigoid, optic neuritis, palindromicrheumatism, PANDAS (Pediatric Autoimmune Neuropsychiatric DisordersAssociated with Streptococcus), paraneoplastic cerebellar degeneration,paroxysmal nocturnal hemoglobinuria (PNH), Parry Romberg syndrome,Parsonnage-Turner syndrome, pars planitis (peripheral uveitis),pemphigus, peripheral neuropathy, perivenous encephalomyelitis,pernicious anemia, POEMS syndrome, polyarteritis nodosa, type I, II, &III autoimmune polyglandular syndromes, polymyalgia rheumatic,polymyositis, postmyocardial infarction syndrome, postpericardiotomysyndrome, progesterone dermatitis, primary biliary cirrhosis, rimarysclerosing cholangitis, psoriasis, psoriatic arthritis, idiopathicpulmonary fibrosis, pyoderma gangrenosum, pure red cell aplasia,Raynauds phenomenon, reactive arthritis, reflex sympathetic dystrophy,Reiter's syndrome, relapsing polychondritis, restless legs syndrome,retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis,sarcoidosis, Schmidt syndrome, scleritis, scleroderma, Sjogren'ssyndrome, sperm & testicular autoimmunity, stiff person syndrome,subacute bacterial endocarditis (SBE), Susac's syndrome, sympatheticophthalmia, Takayasu's arteritis, temporal arteritis/giant cellarteritis, thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome,transverse myelitis, type 1 diabetes, ulcerative colitis,undifferentiated connective tissue disease (UCTD), uveitis, vasculitis,vesiculobullous dermatosis, vitiligo, and Wegener's granulomatosis (nowtermed Granulomatosis with Polyangiitis (GPA).

Any suitable infection may be treated with the antibodies providedherein. Illustrative suitable infections include, for example, hepatitisA virus, hepatitis B virus, hepatitis C virus (HCV), humanimmunodeficiency virus (HIV), and other viral infections.

13. Diagnostic Applications

In some embodiments, the antibodies provided herein are used indiagnostic applications. For example, an ant-PD-1 antibody may be usefulin assays for PD-1 protein. In some aspects the antibody can be used todetect the expression of PD-1 in various cells and tissues. These assaysmay be useful, for example, evaluating cancer and autoimmune disease.

In some diagnostic applications, the antibody may be labeled with adetectable moiety. Suitable detectable moieties include, but are notlimited to radioisotopes, fluorescent labels, and enzyme-substratelabels. In another embodiment of the invention, the anti-PD-1 antibodyneed not be labeled, and the presence thereof can be detected using alabeled antibody which specifically binds to the anti-PD-1 antibody.

14. Affinity Purification Reagents

The antibodies of the invention may be used as affinity purificationagents. In this process, the antibodies may be immobilized on a solidphase such a resin or filter paper, using methods well known in the art.The immobilized antibody is contacted with a sample containing the PD-1protein (or fragment thereof) to be purified, and thereafter the supportis washed with a suitable solvent that will remove substantially all thematerial in the sample except the PD-1 protein, which is bound to theimmobilized antibody. Finally, the support is washed with anothersuitable solvent, such as glycine buffer, pH 5.0, that will release thePD-1 protein from the antibody.

15. Kits

In some embodiments, an anti-PD-1 antibody provided herein is providedin the form of a kit, i.e., a packaged combination of reagents inpredetermined amounts with instructions for performing a procedure. Insome embodiments, the procedure is a diagnostic assay. In otherembodiments, the procedure is a therapeutic procedure.

In some embodiments, the kit further comprises a solvent for thereconstitution of the anti-PD-1 antibody. In some embodiments, theanti-PD-1 antibody is provided in the form of a pharmaceuticalcomposition.

EXAMPLES Example 1: Generation and Primary Screening of Anti-PD-1Antibodies

Antibody Fab or scFv libraries were constructed using a standard overlapextension PCR protocol with mutagenic primers targeting CDRs. SeeHeckman and Pease, Nat. Protoc., 2007, 2:924-932, incorporated byreference in its entirety. Selections for novel antibodies wereperformed using standard ribosome display protocols. See Dreir andPlückthun, Methods Mol. Biol., 2011, Clifton, N.J., 687:283-306,incorporated by reference in its entirety. Specifically, scFv-basedselection formats were performed according to published protocols. SeeHanes and Plückthun, Proc. Natl. Acad. Sci. USA., 1997, 94:4937-4942,incorporated by reference in its entirety. After multiple rounds ofselection, the DNA from RT-PCR output was cloned into an optimizedvector for cell-free expression using standard molecular biologytechniques. See Yin et al., mAbs, 2012, 4:217-225, incorporated byreference in its entirety. All constructs were HIS- and FLAG-tagged tostreamline purification and testing during screening.

Libraries of antibody variants isolated by the selections weretransformed into E. coli and grown on agar plates with antibiotic(kanamycin). Individual colonies were picked and grown in liquid broth(TB+kanamycin), and used as a template for DNA amplification via rollingcircle amplification (RCA). The variants were then expressed in acell-free protein synthesis reaction as described in Zawada et al.(Biotechnol. Bioeng., 2011, 108:1570-1578, incorporated by reference inits entirety).

Briefly, cell-free extracts were treated with 50 μM iodoacetamide for 30minutes at room temperature (RT; 20° C.) and added to a premixcontaining cell-free reaction components (see Groff et al., mAbs, 2014,6:671-678, incorporated by reference in its entirety) and 10% (v/v) RCADNA template (approximately 10 μg/mL DNA) for variants of interest. Cellfree reactions, at a final volume of 604, were incubated at 30° C. for12 h on a shaker at 650 rpm in 96-well plates. Four hundred toone-thousand-five-hundred colonies were screened, depending on thepredicted diversity of the libraries used in the different selectioncampaigns. Following synthesis, each reaction was diluted 1:50 into PBST(PBS at pH 7.4 with 0.2% Tween-20+0.2% BSA) and the variants expressedin each reaction were tested for functional activity via ELISA-basedbinding to recombinant human PD-1 (ACROBiosystems, Inc., Catalog No.PD1-H5221 or SINO Biological Inc. Catalog No. 10377-H08H).

Standard ELISA-based methods were employed. Specifically, 384-wellplates were coated with 2 μg/mL recombinant PD-1 diluted in bicarbonatebuffer, and then blocked with BSA. Antibody variants of interest wereallowed to bind to the PD-1-coated plates, and detected with secondaryantibodies (e.g., HRP-conjugated anti-human Fc or anti-FLAG) and thendetected with chemiluminescent substrate (Pierce ELISA SuperSignal™Substrate). Plates were analyzed on a Molecular Devices SpectraMax® M5plate reader. Top hits were selected based on ELISA signal orsignal/noise ratio and sequenced. Based on functional activity andsequence analysis, a subset of variants was selected for furtherscale-up and characterization.

Example 2: Secondary Screening of Antibody Variants

The top leads from the initial round of screening were cultured andplasmids encoding the antibody genes of interest were isolated using aQIAprep 96 Turbo® Miniprep Kit (Qiagen) according to the manufacturer'sinstructions. DNA was added to 4 mL of cell-free reaction medium toachieve a final concentration of 10 μg/mL. The cell-free reaction mediumwas then incubated overnight for 12 hr at 30° C., at 650 rpm.

The expressed variants from clarified cell-free reactions were purifiedvia immobilized metal ion affinity chromatography (IMAC) using asemi-automated high throughput batch purification method. Briefly,purifications were performed in a 96-well plate format where 50 μL/wellof IMAC resin (Ni Sepharose® High Performance, GE Healthcare) wasequilibrated in IMAC binding buffer (50 mM Tris pH 8.0, 300 mM NaCl, 10mM imidazole), incubated with 1 mL cell-free reaction for 15 minutes,followed by two washes in IMAC binding buffer. His-tagged antibodyvariants were then eluted using 200 μL IMAC elution buffer (50 mM TrispH 8.0, 300 mM NaCl, 500 mM imidazole), and buffer exchanged into PBSusing a 96-well Zeba™ plate (7 kDa MWCO, Thermo Fisher). Purifiedantibodies were quantified via high throughput capillary electrophoresisusing the LabChip GXII® (Perkin Elmer) against a trastuzumab standardcurve, according to the manufacturer's instructions.

Example 3: Hybridoma Generation

Balb/C mice were immunized with the extracellular domain of human PD-1fused with human Fc (R&D Systems, supra) using standard immunizationmethods. The spleens and/or lymph nodes of the mice were harvested andfused with P3X cells to generate the hybridomas (Aragen Biosciences,Morgan Hill, Calif.), similar to what has been previously described. SeeChronopoulou et al., Methods Mol. Biol., 2014, 1131:47-70; and Kim etal., Methods Mol. Biol., 2014, 1131:33-45, each of which is incorporatedby reference in its entirety.

Total RNA was extracted from hybridoma cells using an RNeasy® Mini Kit(Qiagen) and converted to cDNA using a SMARTer™ RACE cDNA AmplificationKit (Clontech). Positive clones were identified by gel electrophoresis,cloned using a TOPO® kit (Invitrogen), and sequenced using standardSanger methods.

Mouse single-chain antibodies were constructed by using total genesynthesis using codons optimized for E. coli. The genes encoding theantibodies were cloned into a standard cell-free expression vector. SeeYin et al., mAbs, 2012, 4:217-225, incorporated by reference in itsentirety.

The CDRs from m1E9 were grafted onto human antibody frameworks VH3-21,VH3-7, Vκ1-9, and Vκ3-11 by standard methodology to yield humanizedantibodies h1E9-1, h1E9-2, h1E9-4, and h1E9-5. See Kuramochi et al.,Methods Mole. Biol., 2014, 1060:123-137, incorporated by reference inits entirety. The same method was used to graft the CDRs from m4B10 ontohuman antibody frameworks VH3-15, Vκ3-11, Vκ3-20, and Vκ4-1 to yieldhumanized antibodies h4B10-1, h4B10-2, and h4B10-3.

Example 4: Kinetic Analysis of Selected Antibody Variants

Human PD-1 (ACROBiosystems, Inc., Catalog No. PD1-H5221), cynomolgusPD-1 (ACROBiosystems, Inc., Catalog No. PD1-H5254), and murine PD-1 (R&DSystems Inc., Catalog No. 1021-PD-100) were used, as indicated, forkinetic analysis.

Monoclonal anti-FLAG M2 IgG (Sigma-Aldrich #F9291) was immobilized ontoa CMS chip (GE Life Sciences) using amine coupling chemistry (from AmineCoupling Kit, GE Life Sciences). The immobilization steps were carriedout at a flow rate of 25 μl/min in 1×HBS-EP+ buffer (GE Life Sciences;10× Stock diluted before use). The sensor surfaces were activated for 7min with a mixture of NHS (0.05 M) and EDC (0.2 M). The Anti-FLAG M2 IgGwas injected over all 4 flow cells at a concentration of 25 μg/ml in 10mM sodium acetate, pH 4.5, for 7 min. Ethanolamine (1 M, pH 8.5) wasinjected for 7 min to block any remaining activated groups. An averageof 12,000 response units (RU) of capture antibody was immobilized oneach flow cell.

Off-rate and kinetic binding experiments were performed at 25° C. using1×HBS-EP+ buffer. Test and control antibodies were injected over theAnti-FLAG surface at concentrations of 5-10 μg/mL for 12 seconds at aflow rate of 10 μl/min on flow cells 2, 3 and 4, followed by a bufferwash for 30 seconds at the same flow rate. Kinetic characterization ofantibody samples was carried out with a single concentration of antigen(for off-rate ranking) or a 1:2 dilution series of antigen (for kineticcharacterization) and 1 injection of 0 nM antigen (i.e., buffer alone).After capturing ligand (anti-PD-1 antibody) on the anti-FLAG surface,the analyte (huPD1-His) was bound at 50, 25, 12.5, 6.25 and 0 nM for 180seconds, followed by a 600 second dissociation phase at a flow rate of50 μl/min. Between each ligand capture and analyte binding cycle,regeneration was carried out using 2 injections of 10 mM glycine pH 2.0for 30 seconds at 30 μL/min, followed by a 30 second buffer wash step.

The data was fit with the Biacore T200 Evaluation software, using a 1:1Langmuir binding model. K_(D) (affinity, nM) was determined as a ratioof the kinetic rate constants calculated from the fits of theassociation and dissociation phases.

Example 5: PD-1-PD-L1 Competition ELISA

Anti-PD1 antibodies were tested for their ability to block a PD-1/PD-L1interaction. PD-1 (ACROBiosystems, Inc.) was adsorbed on 384-well whiteMaxisorp® plates (Nunc) at 2 μg/mL in sodium bicarbonate buffer (pH 8.9)and incubated at 30° C. for 1 hour or overnight at 4° C. The plate waswashed 3 times with PBS pH 7.4 with 0.05% Tween20 and blocked with 2%bovine serum albumin (BSA) in PBS pH 7.4+0.1% Tween20 for 1 hour at 30°C.

The blocking solution was aspirated, and a dilution series of antibodywas mixed with 100 nM PD-L1-Fc (ACROBiosystems, Inc.) in 0.2% BSA in PBSpH 7.4+0.1% Tween20 (diluent buffer) and incubated at 30° C. for 1 hour.The plate was washed, and 10 nM anti-PD-L1 antibody (BioLegend, clone29E.2A3) in diluent buffer was added to all wells. After a 1 hourincubation at 30° C., the plate was washed and incubated withHRP-conjugated anti-mouse Fc (Jackson Laboratories), followed bydetection with SuperSignal™ Pico Chemiluminescent Substrate (ThermoPierce). Luminescence was detected on a SpectraMax® M5 plate reader(Molecular Devices).

Example 6: Cell Binding Experiments

Antibodies with expression levels >250 nM and mouse IgGs from hybridomaswere tested in a fluorescence-activated cell sorting (FACS) cell-bindingassay. Chinese Hamster Ovary (or CHO) Cells stably expressing the humantarget molecule PD-1 on the cell surface (CHO-PD1) were used to screenfor binding. Parental CHO cells were used as a negative control todetermine background-binding levels. Parental CHO cells and CHO-PD1cells were cultured in RPMI w/10% FCS penicillin/streptomycin(Pen/Strep) and glutamine (or Gln) and split every 3-4 days at 10⁵cells/mL.

A mix of parental CHO cells and CHO-PD1 cells was prepared as follows:Parental CHO cells were washed 2× in PBS then incubated in PBScontaining 1 μM CellTrace™ Oregon Green488® (Life Technologies) at 37°C. for 30 minutes. Cells were then washed 2× with RPMI w/10% fetal calfserum (FCS), washed 2× with FACS buffer (PBS w/2% FCS), suspendedthoroughly in ice-cold FACS buffer at a final concentration of 2×10⁶cells/mL and kept on ice. CHO-PD1 cells were similarly washed with FACSbuffer and kept on ice at 2×10⁶ cells/mL. Parental CHO cells and CHO-PD1cells were then mixed to obtain a 1:1 cell suspension and seeded at 100μL per well on 96 well polypropylene plates. Plates were spun at 1500rpm for 5 minutes and cell pellets were suspended in 50 μL FACS buffercontaining 6-12 point dilutions of anti-PD-1 variants starting fromconcentrations of −100-200 nM antibody, dispensed using BioMek FX(Beckman Coulter). Cells were then incubated on ice for 1 hr, washedwith FACS buffer and incubated for 1 hr on ice with 50 μL FACS buffercontaining 2.5 μg/ml R-phycoerythrin-conjugated goat anti-Human IgG(Jackson ImmunoResearch) or AF647-conjugated goat anti-mouse IgG (LifeTechnologies) dispensed using BioMek FX (Beckman Coulter). Cells werethen washed 2× with FACS buffer and fixed for 10 minutes in 200 μL PBSwith 2% paraformaldehyde (PFA) prior to fluorescence detection. Sampleswere acquired using a Becton Dickinson LSRII FACS. Mean FluorescenceIntensity of PD-1 antibody binding was analyzed using FlowJo® software(Tree Star, Inc.).

Example 7: Cell-Based Ligand Competition Experiments

Variants that showed cell-binding activity were tested in afluorescence-activated cell sorting (FACS) cell-based competition assay.CHO cells stably expressing the human target molecule PD-1 on the cellsurface (CHO-PD1) were used to screen for antibodies that compete withhFc-tagged recombinant human PD-L1 or PD-L2 proteins (R&D systems) forbinding to PD-L1 expressed on the cell surface.

CHO-PD1 cells were cultured in RPMI with 10% FCS Pen/Strep and Gln andsplit every 3-4 days at 10⁵ cells/ml. Cells were washed 2× with FACSbuffer (PBS w/2% FCS), thoroughly in ice-cold FACS buffer at a finalconcentration of 1×10⁶ cells/ml and seeded at 1004 per well on 96 wellpolypropylene plates. Plates were spun at 1500 rpm for 5 minutes andcell pellets were suspended in 50 μL FACS buffer containing 8 point 1:3dilutions (2× concentrated) of anti-PD-1 antibody variants, startingfrom high concentration of −200 nM. 50 μL FACS buffer containing a fixedamount of either 6 μg/ml rhPDL2-Fc or 50 μg/ml rhPDL1-Fc proteins werethen added to the cells. Cell were then incubated on ice for 1 hr,washed with FACS buffer and incubated for 1 hr on ice with 50 μl FACSbuffer containing 2.5 μg/ml R-phycoerythrin-conjugated anti-human IgG(Jackson ImmunoResearch). Cells were then washed 2× with FACS buffer andfixed for 10 minutes in 200 μl PBS with 2% PFA prior to acquisition.Samples were acquired using a Becton Dickinson LSRII FACS. MeanFluorescence Intensity of rhPDL1 or rhPDL2 protein binding was analyzedusing FlowJo® software (Tree Star, Inc.).

Example 8: Evaluating the Effect of Anti-PD-1 Antibodies on InterferonGamma Production in a Mixed Lymphocyte Reaction

Anti-PD-1 antibodies were functionally tested for potency in blockingthe PD-1 pathway in a peripheral blood mononuclear cell (PBMC) two-waymixed lymphocyte reaction (MLR) assay by measuring interferon gamma(IFN-g) secretion in cell culture medium. 1×10⁵ human PBMC from 2allogeneic donors were co-cultured in RPMI media+10% FBS in a totalvolume of 150 μl in a 96-well U-bottom plate. Anti-PD-1 antibodies wereadded at specific concentrations to each well. Isotype control antibody,non-PD-1 targeting antibody, or nothing were used as a negativecontrols. Cells were cultured for 5 days at 37° C. At day 5, conditionedmedia was collected and levels of IFN-g were measured using DuoSet®ELISA kits (R&D Systems).

Example 9: Characteristics of Antibodies Isolated from Primary andSecondary Screen

Table 1 shows the characteristics of scFv-Fc antibodies (VH1-18/Vλ3-25)isolated as described in Examples 1-2, and characterized as describedabove.

TABLE 1 Characteristics of antibodies isolated as described in Examples1-2, and characterized as described above. Cell Binding, K_(D) MurineCynomolgus MLR k_(a) (1/Ms) k_(d) (1/s) K_(D) (M) (nM) PD-1 PD-1Activity Human Human Human PD-L1 Human Binding, Binding, K_(D) (IFNgClone ID PD-1 PD-1 PD-1 Competition PD-1 K_(D) (M) (M) release) 1353-A094.88E+05 1.24E−02 2.55E−08 yes, ++ 0.2 Not Not tested Not (SEQ ID NO:238) tested tested 1353-C07 1.23E+06 1.87E−02 1.52E−08 yes 0.4 Not Nottested Not (SEQ ID NO: 239) tested tested 1353-E07 7.37E+05 7.01E−039.52E−09 yes 0.9 Not Not tested Not (SEQ ID NO: 240) tested tested1353-F09 6.87E+05 7.47E−03 1.09E−08 yes 1 Not Not tested Not (SEQ ID NO:241) tested tested 1353-G08 5.63E+05 2.54E−03 4.50E−09 yes, +++ 0.36.09E−08 2.43E−08 Not (SEQ ID NO: 242) tested 1353-G10 5.16E+05 9.80E−041.90E−09 yes, ++ 0.7 6.22E−08 1.55E−08 positive (SEQ ID NO: 243)1353-H08 2.48E+05 1.18E−03 4.76E−09 yes, +++ 0.2 Not Not tested Not (SEQID NO: 244) tested tested 1353-H09 7.98E+05 3.59E−03 4.50E−09 yes, ++0.8 9.08E−09 2.22E−08 positive (SEQ ID NO: 245)

Example 10: Characteristics of Murine Hybridoma Antibodies

Table 2 shows the characteristics of IgG antibodies isolated asdescribed in Example 3, and characterized as described above.

TABLE 2 Characteristics of murine hybridoma antibodies, characterized asdescribed above. PD-L1 PD-L2 Biacore Biacore Cell, Competition,Competition, Human Cynomolgus MLR, EC50¹ IC50² Binding, IC₅₀ ⁴ IC₅₀ ⁴PD1 K_(D) K_(D) IFNg Clone ID (nM) (nM) K_(D) ³ (nM) (nM) (nM) (M) (M)secretion 1B10 1.71 9.47 3.2 5.96 0.56 1.04E−08 2.56E−09 positive VH:SEQ ID NO: 255, with serine prepended to the sequence VL: SEQ ID NO: 2791E9 0.33 0.89 2.9 5.86 0.58 9.90E−09 2.54E−09 positive VH: SEQ ID NO:256 VL: SEQ ID NO: 280 4B10 0.47 1.43 1.39 1.99 0.01 9.13E−10 5.61E−10positive VH: SEQ ID NO: 257 VL: SEQ ID NO: 281 10B4 0.59 1.32 1.34 2.530.18 2.52E−10 1.95E−10 positive VH: SEQ ID NO: 258 VL: SEQ ID NO: 282¹Binding of human PD-1 (ACROBiosystems, Inc., Cat. No. PD1-H5221) viaELISA. ²Competition against PD-L1 (ACROBiosystems, Inc., Cat No.PD1-H5258) ³CHO cell line overexpressing human PD-1. ⁴Cell-basedcompetition. i.e., Inhibition of PD-L1 or PD-L2 binding to CHO cellsoverexpressing human PD-1 is inhibited by IgG.

Example 11: Characteristics of Humanized Antibodies

Table 3 shows the characteristics of humanized scFv antibodies derivedfrom the murine hybridoma antibodies of Example 10, and characterized asdescribed above.

TABLE 3 Characteristics of humanized antibodies, characterized asdescribed above. k_(a) (1/Ms) k_(d) (1/s) K_(D) (M) Clone Human PD-1Human PD-1 Human PD-1 h1E9-1 (VH3-21-Vκ1-9) 1.82E+05 4.68E−04 2.58E−09scFv (SEQ ID NO: 227) h1E9-2 (VH3-21-Vκ3-11) 4.74E+04 1.82E−03 3.85E−08scFv (SEQ ID NO: 228) h1E9-4 (VH3-7-Vκ1-9) 1.85E+05 6.79E−04 3.66E−09scFv (SEQ ID NO: 229) h1E9-5 (VH3-7-Vκ3-11) 2.00E+05 6.28E−04 3.15E−09scFv (SEQ ID NO: 230) m4B10 scFv 8.12E+04 4.17E−04 5.14E−09 (SEQ ID NO:237) h4B10-1 (VH3-15-Vκ3- 1.21E+06 2.99E−03 2.47E−09 11) scFv (SEQ IDNO: 231) h4B10-2 (VH3-15-Vκ3- 1.16E+06 3.24E−03 2.79E−09 20) scFv (SEQID NO: 232) h4B10-3 (VH3-15-Vκ4-1) 5.13E+05 6.17E−04 1.20E−09 scFv (SEQID NO: 233) m1B10 scFv 1.86E+05 2.39E−03 1.28E−08 (SEQ ID NO: 235)

Example 12: Thermal Stability Data

Table 4 provides thermal stability of selected antibodies, as determinedby differential scanning fluorimetry (DSF).

TABLE 4 Thermal stability of selected antibodies, as determined by DSF.Tm1 Tm2 Variant ID Target Scaffold (° C.) (° C.) 1353-G12 PD1 scFv-Fc48.2 1353-G08 PD1 scFv-Fc 45.3 1353-G10 PD1 scFv-Fc 48.6 1353-H09 PD1scFv-Fc 49.4 m1B10 PD1 scFv 55.3 h1E9-1 PD1 scFv 52.5 h1E9-2 PD1 scFv50.9 h1E9-4 PD1 scFv 55.2 h1E9-5 PD1 scFv 51.7 h4b10-1 PD1 scFv 45.559.4 h4b10-2 PD1 scFv 45 61.7 h4b10-3 PD1 scFv 52.1 59.3

Example 13: Construction and Evaluation of h1E9-4 and h1E9-5 IgGs

Variable domains from h1E9-4 scFv (V_(H): SEQ ID NO: 260; V_(L): SEQ IDNO: 284), and h1E9-5 scFv (V_(H): SEQ ID NO: 261; V_(L): SEQ ID NO: 285)were grafted onto human antibody constant domains to generate human IgG1antibodies based on these scFvs.

Specifically, the V_(H) sequences were grafted onto C_(H1)-C_(H2)-C_(H3)constant domains to yield full-length IgG HCs with C-terminal FlagHistags (GSGDYKDDDDKGSGHHHHHH; SEQ ID NO: 294) for ease of purification andassay development. The V_(L) sequences were grafted onto CL domains toyield full-length IgG LCs. The sequence for both the h1E9-4 and h1E9-5HCs, with C-terminal FlagHis tag, is provided in SEQ ID NO: 302. Thesequence for the h1E9-4 LC is provided in SEQ ID NO: 304. The sequencefor the h1E9-5 LC is provided in SEQ ID NO: 303.

The IgGs were expressed in a cell-free reaction, as described in Example1, and purified using the FlagHis tags. The affinity of the IgGs forPD-1 was measured by surface plasmon resonance (Biacore®) and determinedto be essentially equivalent to that of the parent scFvs. Affinity datais provided in Table 5.

TABLE 5 Affinity of h1E9-4 and h1E9-5 IgGs for PD-1 antigen. SampleLigand k_(a) (1/Ms) k_(d) (1/s) K_(D) (M) PD-1- h1E9-4 IgG 3.56E+044.42E−04 1.24E−08 his PD-1- h1E9-5 IgG 2.43E+04 4.71E−04 1.94E−08 his

Example 14: Construction and Evaluation of h1E9 Humanized IgGs

The CDRs for m1E9 were grafted onto human antibody frameworks V_(H)3-23,VH3-30, Vk4-1, Vk3-20, Vk2-28, and Vk1-33 by standard methodology (T.Kuramochi, T. Igawa, H. Tsunoda, and K. Hattori, Method in MolecularBiology, Human Monoclonal Antibodies: Methods and Protocols, 1060,123-137) to yield humanized antibodies h1E9-HC1, h1E9-HC2, h1E9-HC3,h1E9-LC1, h1E9-LC2, h1E9-LC3, and h1E9-LC4. The HC constructs includedC-terminal FlagHis tags (GSGDYKDDDDKGSGHHHHHH; SEQ ID NO: 294) for easeof purification and assay development. The resulting h1E9 heavy chainswere: h1E9-HC3 (SEQ ID NO:324), h1E9-HC2 (SEQ ID NO:325), and h1E9-HC3(SEQ ID NO:326). The resulting light chains were: h1E9-LC4 (SEQ IDNO:327), h1E9-LC3 (SEQ ID NO:328), h1E9-LC2 (SEQ ID NO:329), andh1E9-LC1 (SEQ ID NO:330).

The humanized heavy chains h1E9-HC1, h1E9-HC2, h1E9-HC3, and h1E9-5 werepaired with humanized IgG light chains h1E9-LC1, h1E9-LC2, h1E9-LC3, andh1E9-LC4. These 16 combinations were then assessed for binding to humanPD1 in vitro by surface plasmon resonance (Biacore®) and subsequentlyhuman and cynomolgus PD1 binding on CHO cells using FACS as describedbelow.

Surface plasmon resonance (Biacore data is provided in Table 6.

TABLE 6 Affinity of h1E9 humanized IgGs for PD-1 antigen. Sample LigandSEQ ID NOS K_(D) (M) PD-1-his h1E9-HC1 × h1E9-LC1 326 330 5.08E−09PD-1-his h1E9-HC1 × h1E9-LC2 326 329 1.90E−08 PD-1-his h1E9-HC1 ×h1E9-LC3 326 328 6.38E−09 PD-1-his h1E9-HC1 × h1E9-LC4 326 327 6.33E−09PD-1-his h1E9-HC2 × h1E9-LC1 325 330 3.11E−09 PD-1-his h1E9-HC2 ×h1E9-LC2 325 329 1.23E−08 PD-1-his h1E9-HC2 × h1E9-LC3 325 328 1.09E−08PD-1-his h1E9-HC2 × h1E9-LC4 325 327 4.96E−09 PD-1-his h1E9-HC3 ×h1E9-LC1 324 330 3.60E−09 PD-1-his h1E9-HC3 × h1E9-LC2 324 329 3.19E−09PD-1-his h1E9-HC3 × h1E9-LC3 324 328 1.69E−08 PD-1-his h1E9-HC3 ×h1E9-LC4 324 327 9.40E−09 PD-1-his h1E9-5-IgG-HC-FLAG-HIS × 261 3303.70E−09 h1E9-LC1 PD-1-his h1E9-5-IgG-HC-FLAG-HIS × 261 329 1.62E−08h1E9-LC2 PD-1-his h1E9-5-IgG-HC-FLAG-HIS × 261 328 1.63E−08 h1E9-LC3PD-1-his h1E9-5-IgG-HC-FLAG-HIS × 261 327 7.31E−09 h1E9-LC4

The affinity of the IgGs for cell-surface expressed human PD-1 weremeasured according to Example 6, above. Affinity data is provided inTable 7.

TABLE 7 Affinity of h1E9 humanized IgGs for Cell-Surface Human PD-1antigen (nM). h1E9-HC1 h1E9-HC2 h1E9-HC3 h1E9-5 HC SEQ ID SEQ ID SEQ IDSEQ ID NO: 326 NO: 325 NO: 324 NO: 261 h1E9-LC1 SEQ ID 6 7 8 9 NO: 330h1E9-LC2 SEQ ID 12 11 19 16 NO: 329 h1E9-LC3 SEQ ID 12 9 12 11 NO: 328h1E9-LC4 SEQ ID 8 4 9 14 NO: 327

The affinity of the IgGs for cell-surface expressed cynomolgus PD-1 weremeasured according to Example 6, above. Affinity data is provided inTable 8.

TABLE 8 Affinity of h1E9 humanized IgGs for Cell-Surface Cynomolgus PD-1antigen (nM). h1E9-HC1 h1E9-HC2 h1E9-HC3 h1E9-5 HC SEQ ID SEQ ID SEQ IDSEQ ID NO: 326 NO: 325 NO: 324 NO: 261 h1E9-LC1 SEQ ID 7 6 4 4 NO: 330h1E9-LC2 SEQ ID 19 9 11 11 NO: 329 h1E9-LC3 SEQ ID 15 9 8 9 NO: 328h1E9-LC4 SEQ ID 11 6 5 12 NO: 327

Example 15: In Vivo Efficacy of Anti-PD-1 Antibodies on TumorEstablishment and Growth

MC38 colorectal cancer cells (2×10⁶ cells in 0.1 mL PBS) were implantedsubcutaneously on the right flank of C57BL/6 mice (Charles RiverLaboratories). On day 2 post-cell implantation (day 0), mice weretreated with anti-PD-1 or control antibody at a dose of 5 mg/kgintraperitoneally. Animals were dosed on dosing days 0, 4, 8, 11, and14.

The treatment groups were as follows:

(1) PBS vehicle;

(2) control rat IgG2a, clone 2A3;

(3) anti-PD-1 rat IgG2a, clone RMP1-14;

(4) anti-PD-1 human scFv-Fc, clone PD1-F2 (see SEQ ID NOs: 306 and 293for V_(H) and V_(L) sequences, respectively, and SEQ ID NO: 305 for thescFv-Fc);

(5) anti-PD-1 human scFv-Fc, clone 1353-G08 (see SEQ ID NOs: 251 and 275for V_(H) and V_(L) sequences, respectively, and SEQ ID NO: 242 for thescFv-Fc);

(6) anti-PD-1 human scFv-Fc, clone 1353-G10 (see SEQ ID NOs: 252 and 276for V_(H) and V_(L) sequences, respectively, and SEQ ID NO: 243 for thescFv-Fc); and

(7) anti-PD-1 human IgG, clone PD1-F2v (see SEQ ID NOs: 603 and 293 forV_(H) and V_(L) sequences, respectively, SEQ ID NO: 307 for the HCsequence, and SEQ ID NO: 308 for the LC sequence).

Antibodies (2) and (3) were obtained commercially from Bio X Cell.Antibodies (4)-(6) were produced using the cell-free expression methodsdescribed in Example 1. Antibody 7 was produced by mammalian cellexpression using HEK293 cells and standard techniques.

Tumors were measured using an electronic caliper and tumor volumes werecalculated using the formula, volume=(width²×length)/2. Statisticalanalysis was performed via a two-tailed Mann-Whitney test at day 17post-treatment.

At day 17 post-treatment, all anti-PD-1 antibodies with strong mousePD-1 reactivity (i.e., antibodies RMP1-14, PD1-F2, and 1353-G08)resulted in a significant delay in tumor establishment and growth(p<0.01) compared to the PBS and control rat IgG2a treatment groups. Incontrast, antibody 1353-G10, which binds human PD-1 but has weakerreactivity with mouse PD-1, showed no significant effect on tumorestablishment and growth (p>0.05).

FIG. 2 provides a chart of the tumor volume over the 17-days oftreatment with the various antibodies.

Example 16: Cytomegalovirus (CMV) Recall Assay

Anti-PD-1 antibodies were functionally tested for potency in blockingthe PD-pathway in restimulating peripheral blood mononuclear cells(PBMC) from cytomegalovirus-positive (CMV+) human donors by measuringIFN-g secretion in cell culture medium.

CD14+ monocytes and CD3+ T cells were obtained from peripheral bloodmononuclear cells (PBMC) isolated from CMV+ human donors (AllCells,Alameda, Calif.) using MACS Cell Separation kits (Miltenyi Biotec).

CD14+ monocytes were differentiated into immature dendritic cells (DC)by culturing cells at 1×10⁶ cells/ml for 7 days in presence of GM-CSFand IL-4 (Peprotech) in X-Vivo 15 media (Lonza) containing 2% human ABserum (Sigma-Aldrich), penicillin-streptomycin (Corning Mediatech) andGlutaMAX (Life Technologies). Following differentiation, DCs werematured by culturing in X-Vivo 15+2% human AB serum media at 1×10⁶cells/ml for 2 days in the presence of GM-CSF, IL-4, TNF-α, IL-1b, IL-6(Peprotech) and prostaglandin E2 (Sigma-Aldrich).

To set-up the CMV recall assay, mature DCs were collected and washed.10,000 DCs and 100,000 pan CD3+ T cells were plated per well in a96-well U-bottom plate in a total volume of 100 μl media containingpeptide pools for the CMV IE-1 and CMV pp65 proteins (Miltenyi Biotec).

Anti-PD-1 or control human IgG antibody (50 final volume of 150 μl perwell) were added starting at a final concentration of 400 nM with 5-foldserial dilutions. Cells were co-cultured with peptides and antibodiesfor 5-6 days. Conditioned media was collected and tested for human IFN-glevels by ELISA (BD Biosciences).

As shown in FIG. 3, anti-PD-1 human IgG 1353-G10 (SEQ ID NOs: 252 and276) showed significant fold-increase in IFN-g levels (closed circles).The IFN-g levels were substantial compared to Nivolumab IgG (opencircles).

Example 17: DC/CD4+ T Cell Mixed Lymphocyte Reaction (MLR) Assay

Anti-PD-1 antibodies were functionally tested for potency in blockingthe PD-1 pathway in a peripheral blood mononuclear cell (PBMC) two-waymixed lymphocyte reaction (MLR) assay by measuring interferon gamma(IFN-g) secretion in cell culture medium.

CD14+ monocytes and CD4+ T cells were obtained from PBMC isolated fromhuman donors using MACS Cell Separation kits. CD14+ monocytes weredifferentiated into immature DC by culturing cells at 1×10⁶ cells/mlcell density for 7 days in presence of GM-CSF and IL-4 in RPMI mediacontaining 10% fetal bovine serum, penicillin-streptomycin and GlutaMAX.

Following differentiation, DCs were matured by culturing in RPMI+10% FBSmedia at 1×10⁶ cells/ml cell density for 2 days in the presence ofGM-CSF, IL-4, TNF-a, IL-1b, IL-6 and prostaglandin E2.

To set-up the DC/CD4+ T cell MLR, mature DCs were collected and washed.10,000 DCs and 100,000 CD4+ T cells were plated per well in a 96-wellU-bottom plate in a total volume of 100 μl media. Anti-PD-1 or controlhuman IgG antibody (50 μl, final volume of 150 μl per well) were addedstarting at a final concentration of 400 nM with 5-fold serialdilutions. Cells were co-cultured with peptides and antibodies for 5-6days. Conditioned media was collected and tested for human IFN-g levelsby ELISA.

As shown in FIG. 4, anti-PD-1 human IgG 1353-G10 (SEQ ID NOs: 252 and276) showed significant fold-increase in IFN-g levels (closed circles).The IFN-g levels were substantial compared to Nivolumab IgG (opencircles).

Example 18: Effect of PD-1 Blockade on Graft Versus Host Disease (GVHD)Response

Anti-PD-1 antibodies described herein were evaluated for acceleration ofgraft versus host disease (GVHD) in a mouse model. Since PD-1 negativelyregulates immune response, blockade of PD-1 by an effective antibodyshould accelerate the immune and GVHD response.

40 female NSG mice aged 7 weeks and weighing approximately 19-22 g wereused as human PBMC recipients. Groups were randomized into four groupsof ten. On day 0, all animals received 2×10⁷ human PBMC injected via thetail vein. Prior to PBMC injection, animals were dosed intraperitoneally(IP) with either control anti-GFP, Nivolumab anti-PD-1 antibody, or1353-G10 anti-PD1 antibody (SEQ ID NOs: 252 and 276) as detailed inTable 9. Treatment was administered 2× per week until study conclusion.

TABLE 9 Treatment groups huPBMC Total con- Test Dosing number Groupcentration article Dose frequency of doses Route N 1 2 × 10⁷ Control 102×/week 10 IP 10 Anti-GFP mg/kg 2 2 × 10⁷ Anti-PD1 10 2×/week 10 IP 10Nivolumab mg/kg 3 2 × 10⁷ Anti-PD1 10 2×/week 10 IP 10 1353-G10 mg/kg 42 × 10⁷ Anti-PD1  3 2×/week 10 IP 10 1353-G10 mg/kg

Animals were weighed and observed at least two times per week for 5weeks. Monitoring increased to daily when 10% body weight loss (comparedto initial mouse weight) occurred. Animals were euthanized with CO₂ atthe end of five weeks (end of study), when they exhibited weight lossgreater than 20%, or were unable to right themselves, cold to the touch,and moribund (severe hair ruffling, body weight loss, hunched posture,or decreased activity).

Upon sacrifice (after 5 weeks) or pre-mature euthanasia (due to bodyweight loss or moribundity), the spleen, liver, and serum were collectedfor further analyses. The liver and spleen were placed in formalin for24 hours then transferred to 70% ethanol for storage. The serum wasflash frozen and stored at −80° C. All procedures were conductedaccording to the guidelines of the Sutro Institutional Animal Care andUse Committee (IACUC) and Sutro IACUC protocol SU-001-2013

As shown in FIG. 5, anti-PD-1 treatment with human IgG 1353-G10 (SEQ IDNOs: 252 and 276) showed accelerated mouse morbidity by approximatelytwo weeks (median survival of 13 days for 10 mg/kg 1353-G10 versus 27days for anti-GFP control). The onset of GVHD severity was similar inmice treated with high (10 mg/kg) and low (3 mg/kg) 1353-G10. Bodyweight loss was also faster with both doses of 1353-G10 compared tocontrol and Nivolumab (data not shown). Median survival with both dosesof 1353-G10 was less compared to Nivolumab, indicating greater PD-1blockade potency for 1353-G10 in vivo.

Example 19: Sequences

Table 10 provides sequences referred to herein.

TABLE 10 Sequences. SEQ ID NO Molecule Region Scheme Sequence 1 hPD-1MQIPQAPWPVVWAVLQLGWRPGWFLDSPDR PWNPPTFSPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQPG QDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTERRAE VPTAHPSPSPRPAGQFQTLVVGVVGGLLGSLVLLVWVLAVICSRAARGTIGARRTGQPLK EDPSAVPVFSVDYGELDFQWREKTPEPPVPCVPEQTEYATIVFPSGMGTSSPARRGSADG PRSAQPLRPEDGHCSWPL 2 PD1-17 CDR-H1SGGSIRSTRWWS 3 PD1-28 CDR-H1 SYGIS 4 PD1-33 CDR-H1 SYYIH 5 PD1-35 CDR-H1SGAYYWS 6 PD1-F2 CDR-H1 SSYWMS 7 10B4 CDR-H1 Chothia GYIFSSY 8 1353-A09CDR-H1 Chothia GYRFTWY 9 1353-C07 CDR-H1 Chothia GYRFSTF 10 1353-E07CDR-H1 Chothia GYRFETY 11 1353-F09 CDR-H1 Chothia GYRFRQY 12 1353-G08CDR-H1 Chothia GYRFTRY 13 1353-G10 CDR-H1 Chothia GYRFPHY 14 1353-H08CDR-H1 Chothia GYRFTRQ 15 1353-H09 CDR-H1 Chothia GYRFPHY 16 1B10 CDR-H1Chothia GHSITSDY 17 1E9 CDR-H1 Chothia GFTFSTF 18 4B10 CDR-H1 ChothiaGFTFSTY 19 h1E9-1 CDR-H1 Chothia GFTFSTF 20 h1E9-2 CDR-H1 ChothiaGFTFSTF 21 h1E9-4 CDR-H1 Chothia GFTFSTF 22 h1E9-5 CDR-H1 ChothiaGFTFSTF 23 h4B10-1 CDR-H1 Chothia GFTFSTY 24 h4B10-2 CDR-H1 ChothiaGFTFSTY 25 h4B10-3 CDR-H1 Chothia GFTFSTY 26 PD1-17 CDR-H1 ChothiaGGSIGSGGSIRSTR 27 PD1-28 CDR-H1 Chothia GYRFTSY 28 PD1-33 CDR-H1 ChothiaGYTLTSY 29 PD1-35 CDR-H1 Chothia GGSISSGAY 30 PD1-F2 CDR-H1 ChothiaGFTFSSYWCD 31 10B4 CDR-H1 Kabat SYWIG 32 1353-A09 CDR-H1 Kabat WYGIS 331353-C07 CDR-H1 Kabat TFGIS 34 1353-E07 CDR-H1 Kabat TYGIS 35 1353-F09CDR-H1 Kabat QYGIS 36 1353-G08 CDR-H1 Kabat RYGIS 37 1353-G10 CDR-H1Kabat HYGIS 38 1353-H08 CDR-H1 Kabat RQGIS 39 1353-H09 CDR-H1 KabatHYGIS 40 1B10 CDR-H1 Kabat SDYAWN 41 1E9 CDR-H1 Kabat TFGMS 42 4B10CDR-H1 Kabat TYGMS 43 h1E9-1 CDR-H1 Kabat TFGMS 44 h1E9-2 CDR-H1 KabatTFGMS 45 h1E9-4 CDR-H1 Kabat TFGMS 46 h1E9-5 CDR-H1 Kabat TFGMS 47h4B10-1 CDR-H1 Kabat TYGMS 48 h4B10-2 CDR-H1 Kabat TYGMS 49 h4B10-3CDR-H1 Kabat TYGMS 50 PD1-17 CDR-H1 Kabat SGGSIRSTRWWS 51 PD1-28 CDR-H1Kabat SYGIS 52 PD1-33 CDR-H1 Kabat SYYIH 53 PD1-35 CDR-H1 Kabat SGAYYWS54 PD1-F2 CDR-H1 Kabat SYWCDRMS 55 PD1-17 CDR-H2 EIYHSGSTNYNPSLKS 56PD1-28 CDR-H2 WISAYNGNTNYAQKLQG 57 PD1-33 CDR-H2 IINPRGATISYAQKFQG 58PD1-35 CDR-H2 YIYYNGNTYYNPSLRS 59 PD1-F2 CDR-H2 AISGSGGSTYYADSVKG 6010B4 CDR-H2 Chothia FPGSGS 61 1353-A09 CDR-H2 Chothia SAYNGN 62 1353-C07CDR-H2 Chothia SAYNGN 63 1353-E07 CDR-H2 Chothia SAYNGN 64 1353-F09CDR-H2 Chothia SAYNGN 65 1353-G08 CDR-H2 Chothia SAHNGN 66 1353-G10CDR-H2 Chothia SAYNGN 67 1353-H08 CDR-H2 Chothia SAYNGN 68 1353-H09CDR-H2 Chothia SAYNGN 69 1B10 CDR-H2 Chothia SYSGR 70 1E9 CDR-H2 ChothiaSGGGSD 71 4B10 CDR-H2 Chothia SGGGSN 72 h1E9-1 CDR-H2 Chothia SGGGSD 73h1E9-2 CDR-H2 Chothia SGGGSD 74 h1E9-4 CDR-H2 Chothia SGGGSD 75 h1E9-5CDR-H2 Chothia SGGGSD 76 h4B10-1 CDR-H2 Chothia SGGGSN 77 h4B10-2 CDR-H2Chothia SGGGSN 78 h4B10-3 CDR-H2 Chothia SGGGSN 79 PD1-17 CDR-H2 ChothiaYHSGS 80 PD1-28 CDR-H2 Chothia SAYNGN 81 PD1-33 CDR-H2 Chothia NPRGAT 82PD1-35 CDR-H2 Chothia YYNGN 83 PD1-F2 CDR-H2 Chothia SGSGGS 84 10B4CDR-H2 Kabat KIFPGSGSADYNENFKG 85 1353-A09 CDR-H2 KabatWISAYNGNTNYAQKLQG 86 1353-C07 CDR-H2 Kabat WISAYNGNTNYAQKLQG 87 1353-E07CDR-H2 Kabat WISAYNGNTNYAQKLQG 88 1353-F09 CDR-H2 KabatWISAYNGNTNYAQKLQG 89 1353-G08 CDR-H2 Kabat WVSAHNGNTNYAQKLQG 90 1353-G10CDR-H2 Kabat WISAYNGNTNYAQKLQG 91 1353-H08 CDR-H2 KabatWISAYNGNTKYAQKLQG 92 1353-H09 CDR-H2 Kabat WISAYNGNTNYAQKLQG 93 1B10CDR-H2 Kabat YISYSGRTSYNPSLTS 94 1E9 CDR-H2 Kabat TISGGGSDTYYPDSVQG 954B10 CDR-H2 Kabat TISGGGSNTYYSDSVKG 96 h1E9-1 CDR-H2 KabatTISGGGSDTYYPDSVQG 97 h1E9-2 CDR-H2 Kabat TISGGGSDTYYPDSVQG 98 h1E9-4CDR-H2 Kabat TISGGGSDTYYPDSVQG 99 h1E9-5 CDR-H2 Kabat TISGGGSDTYYPDSVQG100 h4B10-1 CDR-H2 Kabat TISGGGSNTYYSDSVKG 101 h4B10-2 CDR-H2 KabatTISGGGSNTYYSDSVKG 102 h4B10-3 CDR-H2 Kabat TISGGGSNTYYSDSVKG 103 PD1-17CDR-H2 Kabat EIYHSGSTNYNPSLKS 104 PD1-28 CDR-H2 Kabat WISAYNGNTNYAQKLQG105 PD1-33 CDR-H2 Kabat IINPRGATISYAQKFQG 106 PD1-35 CDR-H2 KabatYIYYNGNTYYNPSLRS 107 PD1-F2 CDR-H2 Kabat AISGSGGSTYYADSVKG 108 PD1-17CDR-H3 QDYGDSGDWYFDL 109 PD1-28 CDR-H3 DADYSSGSGY 110 PD1-33 CDR-H3AGIYGFDFDY 111 PD1-35 CDR-H3 ASDYVWGGYRYMDAFDI 112 PD1-F2 CDR-H3ENWGSYFDL 113 10B4 CDR-H3 GYGNYLYFDV 114 1353-A09 CDR-H3 DSEYSSGSGY 1151353-C07 CDR-H3 DVDYSSGSGY 116 1353-E07 CDR-H3 DAEYSLGSGY 117 1353-F09CDR-H3 DAEYGSGSGY 118 1353-G08 CDR-H3 DADYGSGSGY 119 1353-G10 CDR-H3DVDYGTGSGY 120 1353-H08 CDR-H3 DVDYGSGSGY 121 1353-H09 CDR-H3 DAEYGSGSGY122 1B10 CDR-H3 GYALDY 123 1E9 CDR-H3 QGYDVYSWFAY 124 4B10 CDR-H3QRDSAWFAS 125 h1E9-1 CDR-H3 QGYDVYSWFAY 126 h1E9-2 CDR-H3 QGYDVYSWFAY127 h1E9-4 CDR-H3 QGYDVYSWFAY 128 h1E9-5 CDR-H3 QGYDVYSWFAY 129 h4B10-1CDR-H3 QRDSAWFAS 130 h4B10-2 CDR-H3 QRDSAWFAS 131 h4B10-3 CDR-H3QRDSAWFAS 132 PD1-17 CDR-H3 QDYGDSGDWYFDL 133 PD1-28 CDR-H3 DADYSSGSGY134 PD1-33 CDR-H3 AGIYGFDFDY 135 PD1-35 CDR-H3 ASDYVWGGYRYMDAFDI 136PD1-F2 CDR-H3 ENWGSYFDL 137 PD1-17 CDR-L1 TRSSGSIASNSVQ 138 PD1-28CDR-L1 SGDALPKQYAY 139 PD1-33 CDR-L1 TGTSNDVGGYNYVS 140 PD1-35 CDR-L1SGSNSNIGSNSVN 141 PD1-F2 CDR-L1 RASQGISSWLA 142 10B4 CDR-L1 KASQSVSDDVA143 1353-A09 CDR-L1 SGDALTTQYAY 144 1353-C07 CDR-L1 SGDALSEQYAY 1451353-E07 CDR-L1 SGDALPKQYAY 146 1353-F09 CDR-L1 SGDALPKQYAY 147 1353-G08CDR-L1 SGDALPMQYGY 148 1353-G10 CDR-L1 SGDALPKQYAY 149 1353-H08 CDR-L1SGDALPKQYAY 150 1353-H09 CDR-L1 SGDALPKQYAY 151 1B10 CDR-L1 RTSSSVNYMH152 1E9 CDR-L1 RASESVDNSGISFMS 153 4B10 CDR-L1 RASENVDDYGVSFMN 154h1E9-1 CDR-L1 RASESVDNSGISFMS 155 h1E9-2 CDR-L1 RASESVDNSGISFMS 156h1E9-4 CDR-L1 RASESVDNSGISFMS 157 h1E9-5 CDR-L1 RASESVDNSGISFMS 158h4B10-1 CDR-L1 RASENVDDYGVSFMN 159 h4B10-2 CDR-L1 RASENVDDYGVSFMN 160h4B10-3 CDR-L1 RASENVDDYGVSFMN 161 PD1-17 CDR-L1 TRSSGSIASNSVQ 162PD1-28 CDR-L1 SGDALPKQYAY 163 PD1-33 CDR-L1 TGTSNDVGGYNYVS 164 PD1-35CDR-L1 SGSNSNIGSNSVN 165 PD1-F2 CDR-L1 RASQGISSWLA 166 PD1-17 CDR-L2EDNQRPS 167 PD1-28 CDR-L2 KDTERPS 168 PD1-33 CDR-L2 DVTNRPS 169 PD1-35CDR-L2 GNNQRPS 170 PD1-F2 CDR-L2 KASTLES 171 10B4 CDR-L2 YAFKRYI 1721353-A09 CDR-L2 KDTERPS 173 1353-C07 CDR-L2 KDTERPS 174 1353-E07 CDR-L2KDTERPS 175 1353-F09 CDR-L2 KDTERPS 176 1353-G08 CDR-L2 KDTERPS 1771353-G10 CDR-L2 KDTERPS 178 1353-H08 CDR-L2 KDTERPS 179 1353-H09 CDR-L2KDTERPS 180 1B10 CDR-L2 ATSKLAS 181 1E9 CDR-L2 TASNQGS 182 4B10 CDR-L2PASNQGS 183 h1E9-1 CDR-L2 TASNQGS 184 h1E9-2 CDR-L2 TASNQGS 185 h1E9-4CDR-L2 TASNQGS 186 h1E9-5 CDR-L2 TASNQGS 187 h4B10-1 CDR-L2 PASNQGS 188h4B10-2 CDR-L2 PASNQGS 189 h4B10-3 CDR-L2 PASNQGS 190 PD1-17 CDR-L2EDNQRPS 191 PD1-28 CDR-L2 KDTERPS 192 PD1-33 CDR-L2 DVTNRPS 193 PD1-35CDR-L2 GNNQRPS 194 PD1-F2 CDR-L2 KASTLES 195 PD1-17 CDR-L3 QSSDSSAVV 196PD1-28 CDR-L3 QSADNSITYRV 197 PD1-33 CDR-L3 SSYTIVTNFEVL 198 PD1-35CDR-L3 AAWDDSLNGPV 199 PD1-F2 CDR-L3 QQSYSTPWT 200 10B4 CDR-L3 QQNYNSPYT201 1353-A09 CDR-L3 QSADNSITYRV 202 1353-C07 CDR-L3 QSADNSITYRV 2031353-E07 CDR-L3 QSADNSITYRV 204 1353-F09 CDR-L3 QSADNSITYRV 205 1353-G08CDR-L3 QSADNSITYRV 206 1353-G10 CDR-L3 QSADNSITYRV 207 1353-H08 CDR-L3QSADNSITYRV 208 1353-H09 CDR-L3 QSADNSITYRV 209 1B10 CDR-L3 QQWISDPWT210 1E9 CDR-L3 QQSKEVPWT 211 4B10 CDR-L3 QQSKEVPWT 212 h1E9-1 CDR-L3QQSKEVPWT 213 h1E9-2 CDR-L3 QQSKEVPWT 214 h1E9-4 CDR-L3 QQSKEVPWT 215h1E9-5 CDR-L3 QQSKEVPWT 216 h4B10-1 CDR-L3 QQSKEVPWT 217 h4B10-2 CDR-L3QQSKEVPWT 218 h4B10-3 CDR-L3 QQSKEVPWT 219 PD1-17 CDR-L3 QSSDSSAVV 220PD1-28 CDR-L3 QSADNSITYRV 221 PD1-33 CDR-L3 SSYTIVTNFEVL 222 PD1-35CDR-L3 AAWDDSLNGPV 223 PD1-F2 CDR-L3 QQSYSTPWT 224 HCASTKGPSVFPLAPSSKSTSGGTAALGCLVK Constant DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLEPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 225 Kappa LCHMTVAAPSVFIFPPSDEQLKSGTASVVCLL NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC 226 LambdaGQPKAAPSVTLFPPSSEELQANKATLVCLI LD SDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVT HEGSTVEKTVAPTECS 227 h1E9-1 scFvMEVQLVESGGGLVKPGGSLRLSCAASGFTF STFGMSWVRQAPGKGLEWVSTISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAE DTAVYYCARQGYDVYSWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSFLSAS VGDRVTITCRASESVDNSGISFMSWYQQKPGKAPKLLIYTASNQGSGVPSRFSGSGSGTE FTLTISSLQPEDFATYYCQQSKEVPWTFGQGTKVEIKGSGDYKDDDDKGSGHHHHHH 228 h1E9-2 scFvMEVQLVESGGGLVKPGGSLRLSCAASGFTF STFGMSWVRQAPGKGLEWVSTISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAE DTAVYYCARQGYDVYSWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSEIVLTQSPATLSLS PGERATLSCRASESVDNSGISFMSWYQQKPGQAPRLLIYTASNQGSGIPARFSGSGSGTD FTLTISSLEPEDFAVYYCQQSKEVPWTFGQGTKVEIKGSGDYKDDDDKGSGHHHHHH 229 h1E9-4 scFvMEVQLVESGGGLVQPGGSLRLSCAASGFTF STFGMSWVRQAPGKGLEWVATISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAE DTAVYYCARQGYDVYSWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQLTQSPSFLSAS VGDRVTITCRASESVDNSGISFMSWYQQKPGKAPKLLIYTASNQGSGVPSRFSGSGSGTE FTLTISSLQPEDFATYYCQQSKEVPWTFGQGTKVEIKGSGDYKDDDDKGSGHHHHHH 230 h1E9-5 scFvMEVQLVESGGGLVQPGGSLRLSCAASGFTF STFGMSWVRQAPGKGLEWVATISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAE DTAVYYCARQGYDVYSWFAYWGQGTLVTVSSGGGGSGGGGSGGGGSEIVLTQSPATLSLS PGERATLSCRASESVDNSGISFMSWYQQKPGQAPRLLIYTASNQGSGIPARFSGSGSGTD FTLTISSLEPEDFAVYYCQQSKEVPWTFGQGTKVEIKGSGDYKDDDDKGSGHHHHHH 231 h4B10-1 scFvMEVQLVESGGGLVKPGGSLRLSCAASGFTF TSYGMSWVRQAPGKGLEWVATISGGGSNTYYSDSVKGRFTISRDDSKNTLYLQMNSLKTE DTAVYYCARQRDSAWFASWGQGTLVTVSSGGGGSGGGGSGGGGSEIVLTQSPATLSLSPG ERATLSCRASENVDDYGVSFMNWYQQKPGQAPRLLIYPASNQGSGIPARFSGSGSGTDFT LTISSLEPEDFAVYYCQQSKEVPWTFGQGTKVEIKGSGDYKDDDDKGSGHHHHHH 232 h4B10-2 scFvMEVQLVESGGGLVKPGGSLRLSCAASGFTF STYGMSWVRQAPGKGLEWVATISGGGSNTYYSDSVKGRFTISRDDSKNTLYLQMNSLKTE DTAVYYCARQRDSAWFASWGQGTLVTVSSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPG ERATLSCRASENVDDYGVSFMNWYQQKPGQAPRLLIYPASNQGSGIPDRFSGSGSGTDFT LTISRLEPEDFAVYYCQQSKEVPWTFGQGTKVEIKGSGDYKDDDDKGSGHHHHHH 233 h4B10-3 scFvMEVQLVESGGGLVKPGGSLRLSCAASGFTF STYGMSWVRQAPGKGLEWVATISGGGSNTYYSDSVKGRFTISRDDSKNTLYLQMNSLKTE DTAVYYCARQRDSAWFASWGQGTLVTVSSGGGGSGGGGSGGGGSDIVMTQSPDSLAVSLG ERATINCRASENVDDYGVSFMNWYQQKPGQPPKLLIYPASNQGSGVPDRFSGSGSGTDFT LTISSLQAEDVAVYYCQQSKEVPWTFGGGTKLEIKGSGDYKDDDDKGSGHHHHHH 234 m10B4 scFv MQVQLQQSGAELMKPGASVKMSCKTTGYIFSSYWIGWVKQRPGHGLEWIGKIFPGSGSAD YNENFKGKATFTVDTSSNTAYMQLSSLTSEDSAVYYCARGYGNYLYFDVWGAGTTVTVSS GGGGSGGGGSGGGGSNIVMTQTPKFLLVSAGDRITITCKASQSVSDDVAWYQQKPGQSPK LLISYAFKRYIGVPDRFTGSGYGTDFTFTISTVQAEDLAVYFCQQNYNSPYTFGGGTKLE LKGSGDYKDDDDKGSGHHHHHH 235 m1B10 scFvMSDVQLQESGPGLVKPSQSLSLTCTVTGHS ITSDYAWNWIRQFPGNKLEWMGYISYSGRTSYNPSLTSRISITRDTSKNQFFLQLNSVTT EDTATYYCARGYALDYWGQGTSVTVSSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEK VTMTCRTSSSVNYMHWFQQKPGSSPKPWIYATSKLASGVPARFSGSGSGTSYSLTISRVE AEDAATYFCQQWISDPWTFGGGTKLEIKGSGDYKDDDDKGSGHHHHHH 236 m1E9 scFv MEVKLVESGGGLVSPGGSLKLSCAASGFTFSTFGMSWVRQTPEKRLEWVATISGGGSDTY YPDSVQGRFIISRYNAKNNLYLQMNSLRPEDTALYYCARQGYDVYSWFAYWGQGTLVTVS AGGGGSGGGGSGGGGSDIILTQSPASLAVSLGQRAAISCRASESVDNSGISFMSWFQQKP GQPPKLLIYTASNQGSGVPARFSGSGSGTEFSLNIHPMEEDDTAMYFCQQSKEVPWTFGG GTKLEIRGSGDYKDDDDKGSGHHHHHH 237 m4B10scFv MEVKLVESGGGLVKPGGSLKLSCAASGFTF STYGMSWVRQTPEKRLQWVATISGGGSNTYYSDSVKGRFTISRDNAKNNLYLQMSSLRSE DTALYYCARQRDSAWFASWGQGTLVTVSAGGGGSGGGGSGGGGSDIVLTQSPASLAVSLG QRATISCRASENVDDYGVSFMNWFQQKPGQPPKLLIYPASNQGSGVPARFSGSGSGTDFS LNIHPMEEDDTAMYFCQQSKEVPWTEGGGTKLEIKGSGDYKDDDDKGSGHHHHHH 238 1353-A09 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF TWYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDSEYSSGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALTTQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFE LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 239 1353-C07 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF STFGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDVDYSSGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALSEQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 240 1353-E07 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF ETYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDAEYSLGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALPKQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF YLSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 241 1353-F09 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF RQYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDAEYGSGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALPKQYAYWYQQKPGQAPVMVLYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 242 1353-G08 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF TRYGISWVRQAPGQGLEWMGWVSAHNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDADYGSGSGYWGQGTLVTVSSGGGVSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALPMQYGYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 243 1353-G10 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF PHYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDVDYGTGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALPKQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 244 1353-H08 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF TRQGISWVRQAPGQGLEWMGWISAYNGNTKYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDVDYGSGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALPKQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT CPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 245 1353-H09 scFv-FcMEVQLVQSGAEVKKPGASVKVSCKASGYRF PHYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSD DTAVYYCARDAEYGSGSGYWGQGTLVTVSSGGGGSGGGGSGGGGSSYELTQPPSVSVSPG QTARTTCSGDALPKQYAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTIS GVQAEDEADYYCQSADNSITYRVFGGGTKVTVLAAGSDQEPKKLAAGSDQEPKSSDKTHT PCPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHH HH 246 10B4 VHQVQLQQSGAELMKPGASVKMSCKTTGYIFS SYWIGWVKQRPGHGLEWIGKIFPGSGSADYNENFKGKATFTVDTSSNTAYMQLSSLTSED SAVYYCARGYGNYLYFDVWGAGTTVTVSS 2471353-A09 VH EVQLVQSGAEVKKPGASVKVSCKASGYRFTWYGISWVRQAPGQGLEWMGWISAYNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDSEYSSGSGYWGQGTLVTVSS 248 1353-C07 VHEVQLVQSGAEVKKPGASVKVSCKASGYRFS TFGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDVDYSSGSGYWGQGTLVTVSS 2491353-E07 VH EVQLVQSGAEVKKPGASVKVSCKASGYRFETYGISWVRQAPGQGLEWMGWISAYNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDAEYSLGSGYWGQGTLVTVSS 250 1353-F09 VHEVQLVQSGAEVKKPGASVKVSCKASGYRFR QYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDAEYGSGSGYWGQGTLVTVSS 2511353-G08 VH EVQLVQSGAEVKKPGASVKVSCKASGYRFTRYGISWVRQAPGQGLEWMGWVSAHNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDADYGSGSGYWGQGTLVTVSS 252 1353-G10 VHEVQLVQSGAEVKKPGASVKVSCKASGYRFP HYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDVDYGTGSGYWGQGTLVTVSS 2531353-H08 VH EVQLVQSGAEVKKPGASVKVSCKASGYRFTRQGISWVRQAPGQGLEWMGWISAYNGNTKY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDVDYGSGSGYWGQGTLVTVSS 254 1353-H09 VHEVQLVQSGAEVKKPGASVKVSCKASGYRFP HYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDAEYGSGSGYWGQGTLVTVSS 255 1B10 VHDVQLQESGPGLVKPSQSLSLTCTVTGHSIT SDYAWNWIRQFPGNKLEWMGYISYSGRTSYNPSLTSRISITRDTSKNQFFLQLNSVTTED TATYYCARGYALDYWGQGTSVTVSS 256 1E9 VHEVKLVESGGGLVSPGGSLKLSCAASGFTFS TFGMSWVRQTPEKRLEWVATISGGGSDTYYPDSVQGRFIISRYNAKNNLYLQMNSLRPED TALYYCARQGYDVYSWFAYWGQGTLVTVSA 257 4B10VH EVKLVESGGGLVKPGGSLKLSCAASGFTFS TYGMSWVRQTPEKRLQWVATISGGGSNTYYSDSVKGRFTISRDNAKNNLYLQMSSLRSED TALYYCARQRDSAWFASWGQGTLVTVSA 258 h1E9-1VH EVQLVESGGGLVKPGGSLRLSCAASGFTFS TFGMSWVRQAPGKGLEWVSTISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAED TAVYYCARQGYDVYSWFAYWGQGTLVTVSS 259 h1E9-2VH EVQLVESGGGLVKPGGSLRLSCAASGFTFS TFGMSWVRQAPGKGLEWVSTISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAED TAVYYCARQGYDVYSWFAYWGQGTLVTVSS 260 h1E9-4VH EVQLVESGGGLVQPGGSLRLSCAASGFTFS TFGMSWVRQAPGKGLEWVATISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAED TAVYYCARQGYDVYSWFAYWGQGTLVTVSS 261 h1E9-5VH EVQLVESGGGLVQPGGSLRLSCAASGFTFS TFGMSWVRQAPGKGLEWVATISGGGSDTYYPDSVQGRFTISRDNAKNSLYLQMNSLRAED TAVYYCARQGYDVYSWFAYWGQGTLVTVSS 262h4B10-1 VH EVQLVESGGGLVKPGGSLRLSCAASGFTFS TYGMSWVRQAPGKGLEWVATISGGGSNTYYSDSVKGRFTISRDDSKNTLYLQMNSLKTED TAVYYCARQRDSAWFASWGQGTLVTVSS 263 h4B10-2VH EVQLVESGGGLVKPGGSLRLSCAASGFTFS TYGMSWVRQAPGKGLEWVATISGGGSNTYYSDSVKGRFTISRDDSKNTLYLQMNSLKTED TAVYYCARQRDSAWFASWGQGTLVTVSS 264 h4B10-3VH EVQLVESGGGLVKPGGSLRLSCAASGFTFS TYGMSWVRQAPGKGLEWVATISGGGSNTYYSDSVKGRFTISRDDSKNTLYLQMNSLKTED TAVYYCARQRDSAWFASWGQGTLVTVSS 265 PD1-17VH QVQLQESGPGVVKPSGTLSLTCAISGGSIG SGGSIRSTRWWSWVRQSPGKGLEWIGEIYHSGSTNYNPSLKSRVTISLDKSRNHFSLRLN SVTAADTAVYYCARQDYGDSGDWYFDLWGK GTMVTVSS266 PD1-28 VH EVQLVQSGAEVKKPGASVKVSCKASGYRFTSYGISWVRQAPGQGLEWMGWISAYNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDADYSSGSGYWGQGTLVTVSS 267 PD1-33 VHQVQLVQSGAEVKKPGASVRVSCKASGYTLT SYYIHWVRQAPGQGLEWMGIINPRGATISYAQKFQGRVTMTRDTSTSTVYMELRNLKSED TALYYCATAGIYGFDFDYWGRGTLVTVSS 268 PD1-35VH QVQLQESGPGLVKPSQTLSLTCTVSGGSIS SGAYYWSWIRQHPGKGLEWIGYIYYNGNTYYNPSLRSLVTISVDASKNQFSLKLSSVTAA DTAVYYCARASDYVWGGYRYMDAFDIWGRG TLITVSS269 PD1-F2 VH EVQLVQSGGGVVQPGRSLRLSCAASGFTFSSYWCDRMSWVRQAPGKGLEWVSAISGSGGS TYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKENWGSYFDLWGQGTTVTVS S 270 10B4 VLNIVMTQTPKFLLVSAGDRITITCKASQSVS DDVAWYQQKPGQSPKLLISYAFKRYIGVPDRFTGSGYGTDFTFTISTVQAEDLAVYFCQQ NYNSPYTFGGGTKLELKR 271 1353-A09 VLSYELTQPPSVSVSPGQTARITCSGDALTTQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 272 1353-C07 VLSYELTQPPSVSVSPGQTARITCSGDALSEQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 273 1353-E07 VLSYELTQPPSVSVSPGQTARITCSGDALPKQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 274 1353-F09 VLSYELTQPPSVSVSPGQTARITCSGDALPKQ YAYWYQQKPGQAPVMVLYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 275 1353-G08 VLSYELTQPPSVSVSPGQTARITCSGDALPMQ YGYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 276 1353-G10 VLSYELTQPPSVSVSPGQTARITCSGDALPKQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 277 1353-H08 VLSYELTQPPSVSVSPGQTARITCSGDALPKQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 278 1353-H09 VLSYELTQPPSVSVSPGQTARITCSGDALPKQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 279 1B10 VLQIVLSQSPAILSASPGEKVTMTCRTSSSVN YMHWFQQKPGSSPKPWIYATSKLASGVPARFSGSGSGTSYSLTISRVEAEDAATYFCQQW ISDPWTFGGGTKLEIK 280 1E9 VLDIILTQSPASLAVSLGQRAAISCRASESVD NSGISFMSWFQQKPGQPPKLLIYTASNQGSGVPARFSGSGSGTEFSLNIHPMEEDDTAMY FCQQSKEVPWTFGGGTKLEIR 281 4B10 VLDIVLTQSPASLAVSLGQRATISCRASENVD DYGVSFMNWFQQKPGQPPKLLIYPASNQGSGVPARFSGSGSGTDFSLNIHPMEEDDTAMY FCQQSKEVPWTFGGGTKLEIK 282 h1E9-1 VLDIQLTQSPSFLSASVGDRVTITCRASESVD NSGISFMSWYQQKPGKAPKLLIYTASNQGSGVPSRFSGSGSGTEFTLTISSLQPEDFATY YCQQSKEVPWTFGQGTKVEIK 283 h1E9-2 VLEIVLTQSPATLSLSPGERATLSCRASESVD NSGISFMSWYQQKPGQAPRLLIYTASNQGSGIPARFSGSGSGTDFTLTISSLEPEDFAVY YCQQSKEVPWTFGQGTKVEIK 284 h1E9-4 VLDIQLTQSPSFLSASVGDRVTITCRASESVD NSGISFMSWYQQKPGKAPKLLIYTASNQGSGVPSRFSGSGSGTEFTLTISSLQPEDFATY YCQQSKEVPWTFGQGTKVEIK 285 h1E9-5 VLEIVLTQSPATLSLSPGERATLSCRASESVD NSGISFMSWYQQKPGQAPRLLIYTASNQGSGIPARFSGSGSGTDFTLTISSLEPEDFAVY YCQQSKEVPWTFGQGTKVEIK 286 h4B10-1 VLEIVLTQSPATLSLSPGERATLSCRASENVD DYGVSFMNWYQQKPGQAPRLLTYPASNQGSGIPARFSGSGSGTDFTLTISSLEPEDFAVY YCQQSKEVPWTFGQGTKVEIK 287 h4B10-2 VLEIVLTQSPGTLSLSPGERATLSCRASENVD DYGVSFMNWYQQKPGQAPRLLTYPASNQGSGIPDRFSGSGSGTDFTLTISRLEPEDFAVY YCQQSKEVPWTFGQGTKVEIK 288 h4B10-3 VLDIVMTQSPDSLAVSLGERATINCRASENVD DYGVSFMNWYQQKPGQPPKLLTYPASNQGSGVPDRFSGSGSGTDFTLTISSLQAEDVAVY YCQQSKEVPWTFGGGTKLEIK 289 PD1-17 VLNFMLTQPHSVSESPGKTVTISCTRSSGSIA SNSVQWYQQRPGSSPTTVIYEDNQRPSGVPDRFSGSIDSSSNSASLTVSGLKTEDEADYY CQSSDSSAVVFGSGTKLTVL 290 PD1-28 VLSYELTQPPSVSVSPGQTARITCSGDALPKQ YAYWYQQKPGQAPVMVIYKDTERPSGIPERFSGSSSGTKVTLTISGVQAEDEADYYCQSA DNSITYRVFGGGTKVTVL 291 PD1-33 VLQSALTQPASVSGSPGQSITISCTGTSNDVG GYNYVSWYQHHPGKAPKLIIYDVTNRPSGVSDRFSGSKSGNTASLTISGLLAEDEGDYYC SSYTIVTNFEVLEGGGTKLTV 292 PD1-35 VLQSVLTQPPSASGTPGQRVTISCSGSNSNIG SNSVNWYQQLPGTAPKLLIYGNNQRPSGVPDRFSGSKSGTSASLAISGLQSENEADYYCA AWDDSLNGPVFGRGTKVTVL 293 PD1-F2 VLDIVMTQSPSTLSASVGDRVTITCRASQGIS SWLAWYQQKPGRAPKVLIYKASTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ SYSTPWTFGQGTKLEIK 294 FlagHisGSGDYKDDDDKGSGHHHHHH Tag 295 IgG1 Fc AAGSDQEPKSSDKTHTCPPCSAPELLGGSS fromVFLFPPKPKDTLMISRTPEVTCVVVDVSHE scFv-Fc DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPEN NYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGS GDYKDDDDKGSG 296 HCASTKGPSVFPLAPSSKSTSGGTAALGCLVK Constant DYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPS NTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA LPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 297 LC RTVAAPSVFIFPPSDEQLKSGTASVVCLLNConstant NFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 298 Linker GGGGSGGGGSGGGGS 299 murineMWVRQVPWSFTWAVLQLSWQSGWLLEVPNG PD-1 PWRSLTFYPAWLTVSEGANATFTCSLSNWSEDLMLNWNRLSPSNQTEKQAAFCNGLSQPV QDARFQITQLPNRHDEHMNILDTRRNDSGIYLCGAISLHPKAKIEESPGAELVVTERILE TSTRYPSPSPKPEGRFQGMVIGIMSALVGIPVLLLLAWALAVFCSTSMSEARGAGSKDDT LKEEPSAAPVPSVAYEELDFQGREKTPELPTACVHTEYATIVFTEGLGASAMGRRGSADG LQGPRPPRHEDGHCSWPL 300 cynoMWVRQVPWSFTWAVLQLSWQSGWLLEVPNG PD-1 PWRSLTFYPAWLTVSEGANATFTCSLSNWSEDLMLNWNRLSPSNQTEKQAAFCNGLSQPV QDARFQITQLPNRHDEHMNILDTRRNDSGIYLCGAISLHPKAKIEESPGAELVVTERILE TSTRYPSPSPKPEGRFQGMVIGIMSALVGIPVLLLLAWALAVFCSTSMSEARGAGSKDDT LKEEPSAAPVPSVAYEELDFQGREKTPELPTACVHTEYATIVFTEGLGASAMGRRGSADG LQGPRPPRHEDGHCSWPL 301 Linker AAGSDQEPK302 h1E9-4 & HC- MEVQLVESGGGLVQPGGSLRLSCAASGFTF h1E9-5 FlagHisSTFGMSWVRQAPGKGLEWVATISGGGSDTY YPDSVQGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARQGYDVYSWFAYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLEPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHHHH 303 h1E9-5 LC MEIVLTQSPATLSLSPGERATLSCRASESVDNSGISFMSWYQQKPGQAPRLLIYTASNQG SGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSKEVPWTFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC304 h1E9-4 LC MDIQLTQSPSFLSASVGDRVTITCRASESVDNSGISFMSWYQQKPGKAPKLLIYTASNQG SGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQSKEVPWTFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC305 PD1-F2v- scFv-Fc MGAHSEVQLVQSGGGVVQPGRSLRLSCAAS scFvFcFlagGFTFSSYWMSWVRQAPGKGLEWVSAISGSG His GSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKENWGSYFDLWGQGTTVT VSSGGGGSGGGGSGGGGSGVHSDIVMTQSPSTLSASVGDRVTITCRASQGISSWLAWYQQ KPGRAPKVLTYKASTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPWTF GQGTKLEIKAAGSDQEPKSSDKTHTCPPCSAPELLGGSSVFLEPPKPKDTLMISRTPEVT CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGKGSGDYKDDDDKGSGHHHHHH 306 PD1-F2vVH EVQLVQSGGGVVQPGRSLRLSCAASGFTES SYWMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAED TAVYYCAKENWGSYFDLWGQGTTVTVSS 307 PD1-F2vHC GAHSEVQLVQSGGGVVQPGRSLRLSCAASG FTESSYWMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSL RAEDTAVYYCAKENWGSYFDLWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCL VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHK PSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK 308 PD1-F2v LCGVHSDIVMTQSPSTLSASVGDRVTITCRAS QGISSWLAWYQQKPGRAPKVLIYKASTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATY YCQQSYSTPWTFGQGTKLEIKRHMTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC 309 1353-E07- CDR-H3DAEYRLGSGY R5 310 1353-A09- CDR-H3 DSEYRSGSGY R5 311 1353-F09- CDR-H3DAEYRSGSGY R5 312 1353-G08- CDR-H3 DADYRSGSGY R5 313 1353-G10- CDR-H3DVDYRTGSGY R5 314 1353-C07- CDR-H3 DVDYRSGSGY R5 315 1353-H08- CDR-H3DVDYRSGSGY R5 316 1353-E07- VH EVQLVQSGAEVKKPGASVKVSCKASGYRFE R5TYGISWVRQAPGQGLEWMGWISAYNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDAEYRLGSGYWGQGTLVTVSS 317 1353-A09- VHEVQLVQSGAEVKKPGASVKVSCKASGYRFT R5 WYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDSEYRSGSGYWGQGTLVTVSS 3181353-F09- VH EVQLVQSGAEVKKPGASVKVSCKASGYRFR R5QYGISWVRQAPGQGLEWMGWISAYNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDAEYRSGSGYWGQGTLVTVSS 319 1353-G08- VHEVQLVQSGAEVKKPGASVKVSCKASGYRFT R5 RYGISWVRQAPGQGLEWMGWVSAHNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDADYRSGSGYWGQGTLVTVSS 3201353-G10- VH EVQLVQSGAEVKKPGASVKVSCKASGYRFP R5HYGISWVRQAPGQGLEWMGWISAYNGNTNY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDVDYRTGSGYWGQGTLVTVSS 321 1353-C07- VHEVQLVQSGAEVKKPGASVKVSCKASGYRFS R5 TFGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDVDYRSGSGYWGQGTLVTVSS 3221353-H08- VH EVQLVQSGAEVKKPGASVKVSCKASGYRFT R5RQGISWVRQAPGQGLEWMGWISAYNGNTKY AQKLQGRVTMTTDTSTNTAYMELRSLRSDDTAVYYCARDVDYRSGSGYWGQGTLVTVSS 323 1353-H09- VHEVQLVQSGAEVKKPGASVKVSCKASGYRFP R5 HYGISWVRQAPGQGLEWMGWISAYNGNTNYAQKLQGRVTMTTDTSTNTAYMELRSLRSDD TAVYYCARDAEYRSGSGYWGQGTLVTVSS 324H1E9-HC3 HC- MEVQLVESGGGLVQPGGSLRLSCAASGFTF FlagHisSTFGMSWVRQAPGKGLEWVATISGGGSDTY YPDSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARQGYDVYSWFAYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHHHH 325 H1E9-HC2 HC- MQVQLVESGGGVVQPGRSLRLSCAASGFTFFlagHis STFGMSWVRQAPGKGLEWVATISGGGSDTY YPDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARQGYDVYSWFAYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHHHH 326 H1E9-HC1 HC- MEVQLLESGGGLVQPGGSLRLSCAASGFTFFlagHis STFGMSWVRQAPGKGLEWVATISGGGSDTY YPDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARQGYDVYSWFAYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSGDYKDDDDKGSGHHHHHH 327 H1E9-LC4 LC MDIVLTQSPDSLAVSLGERATINCRASESVDNSGISFMSWYQQKPGQPPKLLIYTASNQG SGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSKEVPWTFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC328 H1E9-LC3 LC MEIVLTQSPGTLSLSPGERATLSCRASESVDNSGISFMSWYQQKPGQAPRLLIYTASNQG SGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQSKEVPWTFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC329 H1E9-LC2 LC MDIVLTQSPLSLPVTPGEPASISCRASESVDNSGISFMSWYLQKPGQSPQLLIYTASNQG SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCQQSKEVPWTFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC330 H1E9-LC1 LC MDIQLTQSPSSLSASVGDRVTITCRASESVDNSGISFMSWYQQKPGKAPKLLIYTASNQG SGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQSKEVPWTFGQGTKVEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL SSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC331 CDR-H2 Kabat TISGGGSDTYYPDSVQG

EQUIVALENTS

The disclosure set forth above may encompass multiple distinctinventions with independent utility. Although each of these inventionshas been disclosed in its preferred form(s), the specific embodimentsthereof as disclosed and illustrated herein are not to be considered ina limiting sense, because numerous variations are possible. The subjectmatter of the inventions includes all novel and nonobvious combinationsand subcombinations of the various elements, features, functions, and/orproperties disclosed herein. The following claims particularly point outcertain combinations and subcombinations regarded as novel andnonobvious. Inventions embodied in other combinations andsubcombinations of features, functions, elements, and/or properties maybe claimed in this application, in applications claiming priority fromthis application, or in related applications. Such claims, whetherdirected to a different invention or to the same invention, and whetherbroader, narrower, equal, or different in scope in comparison to theoriginal claims, also are regarded as included within the subject matterof the inventions of the present disclosure.

1.-20. (canceled) 21.-67. (canceled)
 68. A method of treating disease ina subject, comprising administering an effective amount of an antibodyconjugate, or a pharmaceutical composition comprising the same, whereinthe antibody conjugate comprises an antibody or antibody fragment of theIgG class comprising: three heavy chain CDRs of the V_(H) region SEQ IDNO: 252, or a variant thereof, and hree light chain CDRs of the V_(L)region SEQ ID NO: 276, or a variant thereof.
 69. A method of diagnosinga disease in a subject, comprising administering an effective amount ofan antibody conjugate, or a pharmaceutical composition comprising thesame, wherein the antibody conjugate comprises an antibody or antibodyfragment of the IgG class comprising: three heavy chain CDRs of theV_(H) region SEQ ID NO: 252, or a variant thereof, and three light chainCDRs of the V_(L) region SEQ ID NO: 276, or a variant thereof.
 70. Themethod of claim 68, wherein the disease is cancer, an autoimmune diseaseor condition, infection, or any combination thereof.
 71. The method ofclaim 68, wherein the pharmaceutical composition comprises the antibodyor antibody fragment of claim 68 and a pharmaceutically acceptablecarrier.
 72. The method of claim 68, wherein the pharmaceuticalcomposition is substantially pure.
 73. The method of claim 72, whereinthe pharmaceutical composition comprises an antibody that is at least95% by mass of the total antibody or antibody fragment mass of saidcomposition.
 74. The method of claim 68, wherein the pharmaceuticalcomposition is administered intramuscularly, intradermally,intraperitoneally, intravenously, subcutaneously administration, or anycombination thereof.
 75. The method of claim 68, wherein the antibody orantibody fragment comprises: a CDR-H1 comprising SEQ ID NO: 13; a CDR-H2comprising SEQ ID NO: 66; a CDR-H3 comprising SEQ ID NO: 119; a CDR-L1comprising SEQ ID NO: 148; a CDR-L2 comprising SEQ ID NO: 177; and aCDR-L3 comprising SEQ ID NO: 206; or b. a CDR-H1 comprising SEQ ID NO:37; a CDR-H2 comprising SEQ ID NO: 90; a CDR-H3 comprising SEQ ID NO:119; a CDR-L1 comprising SEQ ID NO: 148; a CDR-L2 comprising SEQ ID NO:177; and a CDR-L3 comprising SEQ ID NO:
 206. 76. The method of claim 68,wherein the variant of the V_(H) and V_(L) regions have 20 or feweramino acid substitutions, and wherein the substitutions are conservativeamino acid substitutions.
 77. The method of claim 68, wherein theantibody or antibody fragment further comprises at least one constantregion domain.
 78. The antibody of claim 77, wherein the constant regiondomain comprises a sequence selected from SEQ ID NOs: 224-226 and 297.79. The method of claim 68, wherein the antibody or antibody fragment isa monoclonal antibody.
 80. The method of claim 68, wherein the antibodyor antibody fragment is aglycosylated.
 81. The method of claim 68,wherein the antibody fragment is selected from an Fv fragment, a Fabfragment, a F(ab′)₂ fragment, a Fab′ fragment, an scFv (sFv) fragment,and an scFv-Fc fragment.
 82. The antibody fragment of claim 81, whereinthe antibody fragment is an scFv fragment.
 83. The antibody fragment ofclaim 81, wherein the antibody fragment is an scFv-Fc fragment.
 84. Theantibody fragment of claim 83, wherein the scFv-Fc fragment comprises asequence selected from SEQ ID NO: 243 with AAGSDQEPK (SEQ ID NO: 301)removed from the sequence.
 85. The method of claim 68, wherein theantibody or antibody fragment has a k_(a) of about 4.74×10⁴ M⁻¹×sec⁻¹ toabout 1.23×10⁶ M⁻¹×sec⁻¹ when associating with human PD-1 at atemperature of 25° C.
 86. The method of claim 68, wherein the antibodyor antibody fragment has a k_(d) of about 1.87×10⁻² sec⁻¹ to about4.17×10⁻⁴ sec⁻¹ when dissociating from human PD-1 at a temperature of25° C.
 87. The method of claim 68, wherein the antibody or antibodyfragment has a K_(D) of about 3.85×10⁻⁸ M to about 2.52×10⁻¹⁰ M whenbound to human PD-1 at a temperature of 25° C.
 88. The method of claim68, wherein the antibody or antibody fragment specifically binds one ormore of murine PD-1 and cynomolgus PD-1.